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OPTIMIZATION
OF COMPUTER
NETWORKS –
MODELING AND
ALGORITHMS
OPTIMIZATION
OF COMPUTER
NETWORKS –
MODELING AND
ALGORITHMS
A HANDS-ON APPROACH

Pablo Pavón Mariño

This edition first published 2016
© 2016, John Wiley & Sons, Ltd
Registered office
John Wiley & Sons Ltd, The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, United Kingdom
For details of our global editorial offices, for customer services and for information about how to apply for
permission to reuse the copyright material in this book please see our website at www.wiley.com.
The right of the author to be identified as the author of this work has been asserted in accordance with the Copyright,
Designs and Patents Act 1988.
All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any
form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as permitted by the UK
Copyright, Designs and Patents Act 1988, without the prior permission of the publisher.
Wiley also publishes its books in a variety of electronic formats. Some content that appears in print may not be
available in electronic books.
Designations used by companies to distinguish their products are often claimed as trademarks. All brand names and
product names used in this book are trade names, service marks, trademarks or registered trademarks of their
respective owners. The publisher is not associated with any product or vendor mentioned in this book
Limit of Liability/Disclaimer of Warranty: While the publisher and author have used their best efforts in preparing
this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of
this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose. It is
sold on the understanding that the publisher is not engaged in rendering professional services and neither the
publisher nor the author shall be liable for damages arising herefrom. If professional advice or other expert
assistance is required, the services of a competent professional should be sought.
Library of Congress Cataloging-in-Publication Data
Names: Marino, Pablo Pavon, author.
Title: Optimization of computer networks : modeling and algorithms : a
hands-on approach / Pablo Pavon Marino.
Description: Chichester, West Sussex, United Kingdom : John Wiley & Sons,
Inc., [2016] | Includes bibliographical references and index.
Identifiers: LCCN 2015044522 (print) | LCCN 2016000694 (ebook) | ISBN
9781119013358 (cloth) | ISBN 9781119013334 (ePub) | ISBN 9781119013341
(Adobe PDF)
Subjects: LCSH: Network performance (Telecommunication)–Mathematical models.
| Computer networks–Mathematical models. | Computer algorithms.
Classification: LCC TK5102.83 .M37 2016 (print) | LCC TK5102.83 (ebook) | DDC
004.601–dc23
LC record available at http://lccn.loc.gov/2015044522
A catalogue record for this book is available from the British Library.
Set in 10/12pt, TimesLTStd by SPi Global, Chennai, India.

1 2016
To my sons, Pablo and Guille, and to my wife Victoria,
the smiles of my life.
Contents

About the Author xv

Preface xvii

Acknowledgments xxi

1 Introduction 1
1.1 What is a Communication Network? 1
1.2 Capturing the Random User Behavior 4
1.3 Queueing Theory and Optimization Theory 5
1.4 The Rationale and Organization of this Book 6
1.4.1 Part I: Modeling 6
1.4.2 Part II: Algorithms 7
1.4.3 Basic Optimization Requisites: Appendices I, II, and III 10
1.4.4 Net2Plan Tool: Appendix IV 11

Part I MODELING

2 Definitions and Notation 15

2.1 Notation for Sets, Vectors and Matrices 15
2.1.1 Norm Basics 15
2.1.2 Set Basics 16
2.2 Network Topology 17
2.3 Installed Capacities 19
2.4 Traffic Demands 19
2.4.1 Unicast, Anycast, and Multicast Demands 20
2.4.2 Elastic versus Inelastic Demands 21
2.5 Traffic Routing 21
References 22
viii Contents

3 Performance Metrics in Networks 23

3.1 Introduction 23
3.2 Delay 23
3.2.1 Link Delay 23
3.2.2 End-to-End Delay 27
3.2.3 Average Network Delay 27
3.2.4 Convexity Properties 27
3.3 Blocking Probability 28
3.3.1 Link Blocking Probability 28
3.3.2 Demand and Network Blocking Probability 30
3.3.3 Other Blocking Estimations 31
3.3.4 Convexity Properties 34
3.4 Average Number of Hops 34
3.5 Network Congestion 36
3.6 Network Cost 36
3.7 Network Resilience Metrics 37
3.7.1 Shared Risk Groups 40
3.7.2 Simplified Availability Calculations 41
3.7.3 General Model 41
3.8 Network Utility and Fairness in Resource Allocation 44
3.8.1 Fairness in Resource Allocation 44
3.8.2 Fairness and Utility Functions 45
3.8.3 Convexity Properties 47
3.9 Notes and Sources 47
3.10 Exercises 49
References 51

4 Routing Problems 53
4.1 Introduction 53
4.2 Flow-Path Formulation 54
4.2.1 Optimality Analysis 55
4.2.2 Candidate Path List Pre-Computation 58
4.2.3 Ranking of Paths Elaboration 58
4.2.4 Candidate Path List Augmentation (CPLA) 59
4.3 Flow-Link Formulation 61
4.3.1 Flow Conservation Constraints 62
4.3.2 Obtaining the Routing from xde Variables 63
4.3.3 Optimality Analysis 64
4.4 Destination-Link Formulation 65
4.4.1 Obtaining the Routing Tables from xte Variables 67
4.4.2 Some Properties of the Routing Table Representation 67
4.4.3 Comparing Flow-Based and Destination-Based Routing 71
4.5 Convexity Properties of Performance Metrics 71
4.6 Problem Variants 72
4.6.1 Anycast Routing 72
4.6.2 Multicast Routing 74
Contents ix

4.6.3 Non-Bifurcated Routing 75

4.6.4 Integral Routing 77
4.6.5 Destination-Based Shortest Path Routing 77
4.6.6 SRG-Disjoint 1+1 Dedicated Protection Routing 79
4.6.7 Shared Restoration Routing 80
4.6.8 Multi-Hour Routing 81
4.7 Notes and Sources 83
4.8 Exercises 83
References 86

5 Capacity Assignment Problems 88

5.1 Introduction 88
5.2 Long-Term Capacity Planning Problem Variants 89
5.2.1 Capacity Planning for Concave Costs 89
5.2.2 Capacity Planning with Modular Capacities 94
5.2.3 Multi-Period Capacity Planning 97
5.3 Fast Capacity Allocation Problem Variants: Wireless Networks 98
5.3.1 The Wireless Channel 99
5.3.2 Wireless Networks 100
5.3.3 Modeling Wireless Networks 101
5.4 MAC Design in Hard-Interference Scenarios 104
5.4.1 Optimization in Random Access Networks 105
5.4.2 Optimization in Carrier-Sense Networks 109
5.5 Transmission Power Optimization in Soft Interference Scenarios 113
5.6 Notes and Sources 116
5.7 Exercises 117
References 118

6 Congestion Control Problems 120

6.1 Introduction 120
6.2 NUM Model 121
6.2.1 Utility Functions for Elastic and Inelastic Traffic 121
6.2.2 Fair Congestion Control 122
6.2.3 Optimality Conditions 123
6.3 Case Study: TCP 124
6.3.1 Window-Based Flow Control 125
6.3.2 TCP Reno 126
6.3.3 TCP Vegas 131
6.4 Active Queue Management (AQM) 134
6.4.1 A Simplified Model of the TCP-AQM Interplay 135
6.5 Notes and Sources 136
6.6 Exercises 137
References 139

7 Topology Design Problems 141

7.1 Introduction 141
x Contents

7.2 Node Location Problems 142

7.2.1 Problem Variants 143
7.2.2 Results 144
7.3 Full Topology Design Problems 146
7.3.1 Problem Variants 148
7.3.2 Results 150
7.4 Multilayer Network Design 152
7.5 Notes and Sources 154
7.6 Exercises 154
References 157

Part II ALGORITHMS

8 Gradient Algorithms in Network Design 161

8.1 Introduction 161
8.2 Convergence Rates 163
8.3 Projected Gradient Methods 164
8.3.1 Basic Gradient Projection Algorithm 165
8.3.2 Scaled Projected Gradient Method 165
8.3.3 Singular and Ill-Conditioned Problems 168
8.4 Asynchronous and Distributed Algorithm Implementations 169
8.5 Non-Smooth Functions 172
8.6 Stochastic Gradient Methods 174
8.7 Stopping Criteria 176
8.8 Algorithm Design Hints 177
8.8.1 Dimensioning the Step Size 177
8.8.2 Discrete Step Length 178
8.8.3 Heavy-Ball Methods 179
8.9 Notes and Sources 181
8.10 Exercises 181
References 182

9 Primal Gradient Algorithms 184

9.1 Introduction 184
9.2 Penalty Methods 185
9.2.1 Interior Penalty Methods 185
9.2.2 Exterior Penalty Methods 186
9.3 Adaptive Bifurcated Routing 188
9.3.1 Removing Equality Constraints 189
9.3.2 Optimality and Stability 190
9.3.3 Implementation Example 192
9.4 Congestion Control using Barrier Functions 197
9.4.1 Implementation Example 198
9.4.2 Exterior Penalty 200
Contents xi

9.5 Persistence Probability Adjustment in MAC Protocols 201

9.5.1 Implementation Example 203
9.6 Transmission Power Assignment in Wireless Networks 205
9.6.1 Implementation Example 207
9.7 Notes and Sources 210
9.8 Exercises 211
References 213

10 Dual Gradient Algorithms 214

10.1 Introduction 214
10.2 Adaptive Routing in Data Networks 217
10.2.1 Optimality and Stability 219
10.2.2 Implementation Example 219
10.3 Backpressure (Center-Free) Routing 221
10.3.1 Relation between 𝛾, ΔP, and Average Queue Sizes, Qnd 224
10.3.2 Implementation Example 225
10.4 Congestion Control 228
10.4.1 Optimality and Stability Conditions 229
10.4.2 Implementation Example 230
10.5 Decentralized Optimization of CSMA Window Sizes 231
10.5.1 Implementation Example 234
10.6 Notes and Sources 236
10.7 Exercises 236
References 238

11 Decomposition Techniques 240

11.1 Introduction 240
11.2 Theoretical Fundamentals 241
11.2.1 Primal Decomposition 241
11.2.2 Dual Decomposition 244
11.2.3 Other Decompositions 246
11.3 Cross-Layer Congestion Control and QoS Capacity Allocation 247
11.3.1 Implementation Example 249
11.4 Cross-Layer Congestion Control and Backpressure Routing 249
11.4.1 Implementation Example 252
11.5 Cross-Layer Congestion Control and Power Allocation 253
11.5.1 Implementation Example 254
11.6 Multidomain Routing 256
11.6.1 Implementation Example 258
11.7 Dual Decomposition in Non-Convex Problems 259
11.7.1 Implementation Example 261
11.8 Notes and Sources 261
11.9 Exercises 263
References 265
xii Contents

12 Heuristic Algorithms 266

12.1 Introduction 266
12.1.1 What Complexity Theory Tells Us that We cannot Do 266
12.1.2 Our Options 267
12.1.3 Organization and Rationale of this Chapter 268
12.2 Heuristic Design Keys 270
12.2.1 Heuristic Types 270
12.2.2 Intensification versus Diversification 271
12.2.3 How to Assess the Solution Quality 271
12.2.4 Stop Conditions 272
12.2.5 Defining the Cost or Fitness Function 272
12.2.6 Coding the Solution 273
12.3 Local Search Algorithms 273
12.3.1 Design Hints 274
12.4 Simulated Annealing 276
12.4.1 Design hints 277
12.5 Tabu Search 278
12.5.1 Design Hints 280
12.6 Greedy Algorithms 281
12.7 GRASP 282
12.8 Ant Colony Optimization 283
12.8.1 Design Hints 286
12.9 Evolutionary Algorithms 288
12.9.1 Design Hints 289
12.10 Case Study: Greenfield Plan with Recovery Schemes Comparison 291
12.10.1 Case Study Description 291
12.10.2 Algorithm Description 293
12.10.3 Combining Heuristics and ILPs 295
12.10.4 Results 296
12.11 Notes and Sources 297
12.12 Exercises 297
References 299

A Convex Sets. Convex Functions 301

A.1 Convex Sets 301
A.2 Convex and Concave Functions 303
A.2.1 Convexity in Differentiable Functions 303
A.2.2 Strong Convexity/Concavity 306
A.2.3 Convexity in Non-Differentiable Functions 306
A.2.4 Determining the Curvature of a Function 307
A.2.5 Sub-level Sets 310
A.2.6 Epigraphs 311
A.3 Notes and Sources 311
Reference 312
Contents xiii

B Mathematical Optimization Basics 313

B.1 Optimization Problems 313
B.2 A Classification of Optimization Problems 315
B.2.1 Linear Programming 315
B.2.2 Convex Programs 318
B.2.3 Nonlinear Programs 320
B.2.4 Integer Programs 321
B.3 Duality 324
B.3.1 Dual Function 324
B.4 Optimality Conditions 330
B.4.1 Optimality Conditions in Problems with Strong Duality 330
B.4.2 Graphical Interpretation of KKT Conditions 333
B.4.3 Optimality Conditions in Problems Without Strong Duality 336
B.5 Sensitivity Analysis 337
B.6 Notes and Sources 339
References 340

C Complexity Theory 341

C.1 Introduction 341
C.2 Deterministic Machines and Deterministic Algorithms 342
C.2.1 Complexity of a Deterministic Algorithm 342
C.2.2 Worst-Case Algorithm Complexity 343
C.2.3 Asymptotic Algorithm Complexity 343
C.2.4 Complexity is a Real Barrier 345
C.3 Non-Deterministic Machines and Non-Deterministic Algorithms 346
C.3.1 Complexity of a Non-Deterministic Algorithm 347
C.4  and   Complexity Classes 347
C.5 Polynomial Reductions 349
C.5.1 A Polynomial Time Reduction Example 350
C.6  -Completeness 351
C.6.1 An Example Proving  -Completeness for a Problem 352
C.7 Optimization Problems and Approximation Schemes 352
C.7.1 The   Class 353
C.7.2 Approximation Algorithms 354
C.7.3 PTAS Reductions 356
C.7.4  -Complete Problems 356
C.8 Complexity of Network Design Problems 357
C.9 Notes and Sources 357
References 358

D Net2Plan 359
D.1 Net2Plan 359
D.2 On the Role of Net2Plan in this Book 360

Index 363
About the Author

Pablo Pavón Mariño is Associate Professor at the Universidad Politécnica de Cartagena (Spain)
and Head of GIRTEL research group, MSc and Ph.D in Telecommunications, and MSc in
Mathematics, with specialization in operations research. His research interests in the last 15
years are in optimization, planning, and performance evaluation of computer networks. He
has more than a decade track as a lecturer in network optimization courses. He is author or
co-author of more than 100 research papers in the field, published in top journals and inter-
national conferences, as well as several patents. He leads the Net2Plan open-source initiative,
which includes the Net2Plan tool and its associated public repository of algorithms and net-
work optimization resources (www.net2plan.com). Pablo Pavón has served as chair in
international conferences like IEEE HPSR 2011, ICTON 2013 or ONDM 2016. He is Tech-
nical Editor of the Optical Switching and Networking journal, and has participated as Guest
Editor in other journals such as Computer Networks, Photonic Network Communications, and
IEEE/OSA Journal of Optical Communications and Networking.
Other documents randomly have
different content
by employing different kinds of sealing-wax; but is very liable to
“chip” and leave the glass.
Black Varnish is readily prepared by adding a small quantity of
lamp-black to gold-size and mixing intimately. Dr. Carpenter
recommends this as a good finishing varnish, drying quickly and
being free from that brittleness which renders some of the others
almost worthless; but it should not be used in the first process when
mounting objects in fluid.
Amongst these different cements and varnishes I worked a long
time without coming to any decision as to their comparative
qualities, though making innumerable experiments. The harder kinds
were continually cracking, and the softer possessed but little
adhesive power. To find hardness and adhesiveness united was my
object, and the following possesses these qualities in a great
degree:—

India-rubber ½ drachm.
Asphaltum 4 oz.
Mineral naphtha 10 ”

Dissolve the india-rubber in the naphtha, then add the asphaltum—if

necessary heat must be employed.

This is often used by photographers as a black varnish for glass,

and never cracks, whilst it is very adhesive. Dr. Carpenter, however,
states that his experience has not been favourable to it; but I have
used it in great quantities and have never found it to leave the glass
in a single instance when used in the above proportions. The
objections to it are, however, I think easily explained, when it is
known that there are many kinds of pitch, &c., from coal, sold by the
name of asphaltum, some of which are worthless in making a
microscopic cement. When used for this purpose, the asphaltum
must be genuine and of the best quality that can be bought. The
above mixture serves a double purpose—to unite the cell to the
slide, and also as a “finishing” varnish. But it is perhaps more
convenient to have two bottles of this cement, one of which is
thicker than common varnish, to use for uniting the cell, &c.; the
other liquid enough to flow readily, which may be employed as a
surface varnish in finishing the slides.
The brushes or camel-hair pencils should always be cleaned
after use; but with the asphalt varnish above mentioned it is
sufficient to wipe off as carefully as possible the superfluous quantity
which adheres to the pencil, as, when again made use of, the
varnish will readily soften it; but, of course, it will be necessary to
keep separate brushes for certain purposes.
Here it may be observed that every object should be labelled
with name and any other descriptive item as soon as mounted.
There are many little differences in the methods of doing this. Some
write with a diamond upon the slide itself; but this has the
disadvantage of being not so easily seen. For this reason a small
piece of paper is usually affixed to one end of the slide, on which is
written what is required. These labels may be bought of different
colours and designs; but the most simple are quite as good, and
very readily procured. Take a sheet of thin writing paper and brush
over one side a strong solution of gum, with the addition of a few
drops of glycerine as above recommended; allow this to dry, and
then with a common gun-punch stamp out the circles, which may be
affixed to the slides by simply damping the gummed surface, taking
care to write the required name, &c., upon it before damping it, or
else allowing it to become perfectly dry first.
There is one difficulty which a beginner often experiences in
sorting and mounting certain specimens under the microscope, viz.,
the inversion of the objects; and it is often stated to be almost
impossible to work without an erector. But this difficulty soon
vanishes, the young student becoming used to working what at first
seems in contradiction to his sight.
Let it be understood, that in giving the description of those
articles which are usually esteemed necessary in the various parts of
microscopic manipulation, I do not mean to say that without many
of these no work of any value can be done. There are, as all will
allow, certain forms of apparatus which aid the operator
considerably; but the cost may be too great for him. A little thought,
however, will frequently overcome this difficulty, by enabling him to
make, or get made, for himself, at a comparatively light expense,
something which will accomplish all he desires. As an example of
this, a friend of mine made what he terms his “universal stand,” to
carry various condensers, &c., &c., in the following way:—Take a
steel or brass wire, three-sixteenths or one-quarter inch thick and six
or eight inches long; “tap” into a solid, or make rough and fasten
with melted lead into a hollow, ball. (The foot of a cabinet or work-
box answers the purpose very well.) In the centre of a round piece
of tough board, three inches in diameter, make a hemispherical
cavity to fit half of the ball, and bore a hole through from the middle
of this cavity, to allow the wire to pass. Take another piece of board,
about four inches in diameter, either round or square, and one-and-
a-half or two inches thick, make a similar cavity in its centre to
receive the other half of the ball, but only so deep as to allow the
ball to fit tightly when the two pieces of board are screwed together,
which last operation must be done with three or four screws. Let the
hole for the wire in the upper part be made conical (base upwards),
and so large as only to prevent the ball from escaping from its
socket, in order that the shaft may move about as freely as possible.
Turn a cavity, or make holes, in the bottom of the under piece, and
fill with lead to give weight and steadiness. This, painted green
bronze and varnished, looks neat; and by having pieces of gutta-
percha tubing to fit the shaft, a great variety of apparatus may be
attached to it.
Again, a “condenser” is often required for the illumination of
opaque objects. My ingenious friend uses an “engraver’s bottle”
(price 6d.), fills it with water, and suspends it betwixt the light and
the object. Where the light is very yellow, he tints the water with
indigo, and so removes the objectionable colour.
 I merely mention these as examples of what may be done by a
little thoughtful contrivance, and to remove the idea that nothing is
of much value save that which is the work of professional workmen,
and consequently expensive.
 C H A P T E R I I.
TO PREPARE AND MOUNT OBJECTS “DRY.”

The term “dry” is used when the object to be mounted is not

immersed in any liquid or medium, but preserved in its natural state,
unless it requires cleaning and drying.
I have before stated that thorough cleanliness is necessary in
the mounting of all microscopic objects. I may here add that almost
every kind of substance used by the microscopist suffers from
careless handling. Many leaves with fine hairs are robbed of half
their beauty, or the hairs, perhaps, forced into totally different
shapes and groups; many insects lose their scales, which constitute
their chief value to the microscopist; even the glass itself distinctly
shows the marks of the fingers if left uncleaned. Every object must
also be thoroughly dry, otherwise dampness will arise and become
condensed in small drops upon the inner surface of the thin glass
cover. This defect is frequently met with in slides which have been
mounted quickly; the objects not being thoroughly dry when
enclosed in the cell. Many of the cheap slides are thus rendered
worthless. Even with every care it is not possible to get rid of this
annoyance occasionally.
For the purpose of mounting opaque objects “dry,” discs were at
one time very commonly made use of. These are circular pieces of
cork, leather, or other soft substance, from one-quarter to half inch
in diameter, blackened with varnish or covered with black paper, on
which the object is fixed by gum or some other adhesive substance.
They are usually pierced longitudinally by a strong pin, which serves
for the forceps to lay hold of when being placed under the
microscope for examination. Sometimes objects are affixed to both
sides of the disc, which is readily turned when under the object-
glass. The advantage of this method of mounting is the ease with
which the disc may be moved, and so present every part of the
object to the eye save that by which it is fastened to the disc. On
this account it is often made use of when some particular subject is
undergoing investigation, as a number of specimens may be placed
upon the discs with very little labour, displaying all the parts. But
where exposure to the atmosphere or small particles of dust will
injure an object, no advantage which the discs may possess should
be considered, and an ordinary covered cell should be employed.
Small pill-boxes have been used, to the bottom of which a piece of
cork has been glued to afford a ground for the pin or other mode of
attachment; but this is liable to some of the same faults as the disc,
and it would be unwise to use these for permanent objects.
Messrs. Smith and Beck have lately invented, and are now
making, a beautiful small apparatus, by means of which the disc
supporting the object can be worked with little or no trouble into any
position that may prove most convenient, whilst a perforated
cylinder serves for the reception of the discs when out of use, and
fits into a case to protect them from dust. A pair of forceps is made
for the express purpose of removing them from the case and placing
them in the holder.
All dry objects, however, which are to be preserved should be
mounted on glass slides in one of the cells (described in Chapter I.)
best suited to them. Where the object is to be free from pressure,
care must be taken that the cell is deep enough to ensure this.
When the depth required is but small, it is often sufficient to omit
the card, leather, or other circles, and with the “turn-table” before
described, by means of a thick varnish and camel-hair pencil, to
form a ring of the desired depth; but should the varnish not be of
sufficient substance to give such “walls” at once, the first application
may be allowed to dry, and a second made upon it. A number of
these may be prepared at the same time, and laid by for use. When
liquids are used (see Chapter IV.), Dr. Carpenter recommends gold-
size as a good varnish for the purpose, and this may be used in
“dry” mountings also. I have used the asphaltum and india-rubber
(mentioned in Chapter I.), and found it to be everything I could
wish. The cells, however, must be thoroughly dry, and when they
will bear the heat they should be baked for an hour at least in a
tolerably cool oven, by which treatment the latter becomes a first-
rate medium. All dry objects which will not bear pressure must be
firmly fastened to the slide, otherwise the necessary movements
very often injure them, by destroying the fine hairs, &c. For this
purpose thin varnishes are often used, and will serve well enough for
large objects, but many smaller ones are lost by adopting this plan,
as for a time, which may be deemed long enough to harden the
varnish, they exhibit no defect, but in a while a “wall” of the plastic
gum gathers around them, which refracts the light, and thus leads
the student to false conclusions. In all finer work, where it is
necessary to use any method of fixing them to the slide, a solution
of common gum, with the addition of a few drops of glycerine
(Chapter I.), will be found to serve the purpose perfectly. It must,
however, be carefully filtered through blotting paper, otherwise the
minute particles in the solution interfere with the object, giving the
slide a dusty appearance when under the microscope.
When mounting an object in any of these cells, the glass must
be thoroughly cleaned, which may be done with a cambric
handkerchief, after the washing mentioned in Chapter I. If the object
be large, the point of a fine camel-hair pencil should be dipped into
the gum solution, and a minute quantity of the liquid deposited in
the cell where the object is to be placed, but not to cover a greater
surface than the object will totally hide from sight. This drop of gum
must be allowed to dry, which will take a few minutes. Breathe then
upon it two or three times, holding the slide not far from the mouth,
which will render the surface adhesive. Then draw a camel-hair
pencil through the lips, so as to moisten it slightly (when anything
small will adhere to it quite firmly enough), touch the object and
place it upon the gum in the desired position. This must be done
immediately to ensure perfect stability, otherwise the gum will
become at least partially dry and only retain the object imperfectly.
When, however, the objects are so minute that it would be
impossible to deposit atoms of gum small enough for each one to
cover, a different method of proceeding must be adopted. In this
case a small portion of the same gum solution should be placed
upon the slide, and by means of any small instrument—a long
needle will serve the purpose very well—spread over the surface
which will be required. The quantity thus extended will be very
small, but by breathing upon it may be prevented drying whilst being
dispersed. This, like the forementioned, should be then allowed to
dry; and whilst the objects are being placed on the prepared
surface, breathing upon it as before will restore the power of
adherence.
When gum or other liquid cement has been used to fix the
objects to the glass, the thin covers must not be applied until the
slide has been thoroughly dried, and all fear of dampness arising
from the use of the solution done away with. Warmth may be safely
applied for the purpose, as objects fastened by this method are
seldom, if ever, found to be loosened by it. As objects are met with
of every thickness, the cells will be required of different depths.
There is no difficulty in accommodating ourselves in this—the deeper
cells may be readily cut out of thick leather, card, or other substance
preferred (as mentioned in Chapter I.). Cardboard is easily procured
of almost any thickness; but sometimes it is convenient to find a
thinner substance even than this. When thin glass is laid upon a
drop of any liquid upon a slide, every one must have observed how
readily the liquid spreads betwixt the two: just so when any thin
varnish is used to surround an object of little substance, excessive
care is needed lest the varnish should extend betwixt the cover and
slide, and so render it worthless. The slightest wall, however,
prevents this from taking place, so that a ring of common paper may
be used, and serve a double purpose where the objects require no
deeper cell than this forms.
 Many objects, however, are of such tenuity—as the leaves of
many mosses, some of the Diatomaceæ, scales of insects, &c.—that
no cell is requisite excepting that which is necessarily formed by the
medium used to attach the thin glass cover to the slide; and where
the slide is covered by the ornamental papers mentioned in Chapter
I., and pressure does not injure the object, even this is omitted, the
thin glass being kept in position by the cover; but slides mounted in
this manner are frequently injured by dampness, which soon
condenses upon the inner surfaces and interferes both with the
object and the clearness of its appearance.
The thin glass, then, is to be united to the slide, so as to form a
perfect protection from dust, dampness, or other injurious matter,
and yet allow a thoroughly distinct view of the object. This is to be
done by applying to the glass slide round the object some adhesive
substance, and with the forceps placing the thin glass cover (quite
dry and clean) upon it. A gentle pressure round the edge will then
ensure a perfect adhesion, and with ordinary care there will be little
or no danger of breakage. For this purpose gold-size is frequently
used. The asphalt and india-rubber varnish also will be found both
durable and serviceable. Whatever cement may be used, it is well to
allow it to become in some measure “fixed” and dried; but where no
cell or “wall” is upon the slide, this is quite necessary, otherwise the
varnish will be almost certain to extend, as before mentioned, and
ruin the object. It may be stated here that gold-size differs greatly in
its drying powers, according to its age, mode of preparation, &c.
(Chapter IV.)
Should any object be enclosed which requires to be kept flat
during the drying of the cement, it will be necessary to use some of
the contrivances mentioned in Chapter I.
When the slide is thus far advanced, there remains the
“finishing” only. Should the student, however, have no time to
complete his work at once, he may safely leave it at this stage until
he has a number of slides which he may finish at the same time.
There are different methods of doing this, some of which may be
here described.
If ornamental papers are preferred, a small circle must be cut
out from the centre a little less than the thin glass which covers the
object. Another piece of coloured paper is made of the same size,
and a similar circle taken from its centre also, or both may be cut at
the same time. The slide is then covered round the edges with paper
of any plain colour, so that it may extend about one-eighth of an
inch over the glass on every side. The ornamental paper is then
pasted on the “object” surface of the glass, so that the circle shows
the object as nearly in the centre as possible, and covers the edges
of the thin glass. The other coloured paper is then affixed
underneath with the circle coinciding with that above. And here I
may observe, that when this method is used there is no necessity for
the edges of the slide to be “ground,” as all danger of scratching,
&c., is done away with by the paper cover.
Many now use paper covers, about one and a half inches long,
on the upper side of the slide only, with the centre cut out as before,
with no other purpose than that of hiding the edge of the thin glass
where it is united to the slide.
The method of “finishing,” however, which is mostly used at the
present time, is to lay a coating of varnish upon the edge of the thin
glass, and extend it some little way on the slide. When a black circle
is required, nothing serves the purpose better than the gold-size and
lamp-black, or the asphalt and india-rubber varnish, neither of which
is liable to chip; but when used for this, the latter should be rather
thinner, as before advised. Some of these varnishes are preferred of
different colours, which may be made by using the different kinds of
sealing-wax, as described in Chapter I.; but they are always liable to
the defects there mentioned. This circle cannot be made in any
other way but by one of those contrivances which have now centred
in Shadbolt’s turntable. A very little practice will enable the young
student to place his slide so that the circle may be uniform with the
edge of the thin glass.
 The slide is now complete, except the addition of the name and
any other particulars which may be desirable. For this purpose one
of the methods described in Chapter I. must be employed.
Amongst the various classes of microscopic objects now
receiving general attention, the Diatomaceæ may be placed in a
prominent position. They afford endless opportunities of research,
and some very elaborate works have already been issued concerning
them. Professor Smith’s may be mentioned as one containing,
perhaps, the best illustrations. The young student may wish to know
what a diatom is. The “Micrographic Dictionary” gives the following
definition:—“A family of confervoid Algæ, of very peculiar character,
consisting of microscopic brittle organisms.” They are now looked
upon by almost all of our scientific men as belonging to the
vegetable kingdom, though some few still assign them to the animal.
They are almost invariably exceedingly small, so that the unaided
eye can perceive nothing on a prepared slide of these organisms but
minute dust. Each separate portion, which is usually seen when
mounted, is termed a “frustule,” or “testule:” this consists of two
similar parts, composed of silica, between and sometimes around
which is a mass of viscid matter called the “endochrome.” They are
found in almost every description of water, according to the variety:
some prefer sea-water, others fresh, and many are seen nowhere
but in that which is a mixture of both, as the mouths of rivers, &c.
Ditches, ponds, cisterns, and indeed almost every reservoir, yield
abundance of these forms. They are not, however, confined to
“present” life; but, owing to the almost indestructible nature of their
siliceous covering, they are found in a fossil state in certain earths in
great abundance, and are often termed “fossil Infusoria.” Upon these
frustules are generally to be seen lines, or “markings,” of different
degrees of minuteness, the delicacy of which often serves the
purpose of testing the defining power of the object-glasses. Some of
the frustules are triangular, others circular, and, indeed, of almost
every conceivable shape, many of them presenting us with
exquisitely beautiful designs.
 The markings, however, are seldom seen well, if at all, until the
frustules are properly prepared, the different methods of
accomplishing which will be given a little further on.
The collection of the fresh diatoms is so closely connected with
their preservation, that a few notes may be given upon it before we
pass on. For this purpose a number of small bottles must be
provided, which may be placed in a tin box, with a separate
apartment for each, so that all chance of breakage may be done
away with. The diatoms are generally of a light brown colour; and
where they are observed in the water, the bottle may be so placed,
with the mouth closed by the finger, that when the finger is
withdrawn the water will rush in, carrying the diatoms also. If they
are seen upon plants, stones, or any other substance, they may
generally be detached and placed in the bottle. When there is a
green covering upon the surface of the water, a great quantity of
diatoms is usually found amongst it; as also upon the surface of the
mud in those ponds where they abound. In these cases, a broad flat
spoon will be found very useful, and one is now made with a
covering upon the broader portion of it to protect the enclosed
matter from being so readily carried off whilst bringing it to the
surface again. Where there is any depth of water, and the spoon will
not reach the surface of the mud, the bottle must be united to a
long rod, and being then carried through the upper portion with the
mouth downwards, no water will be received into it; but on reaching
the spot required, the bottle-mouth may be turned up, and thus
become filled with what is nearest.
From the stomachs of common fish—as the cod, sole, haddock,
&c.—many specimens of Diatomaceæ may be obtained, but
especially from the crab, oyster, mussel, and other shell-fish.
Professor Smith states that from these curious receptacles he has
taken some with which he has not elsewhere met. To remove them
from any of the small shell-fish, it is necessary to take the fish or
stomach from the shell, and immerse it in strong hot acid (nitric is
the best) until the animal matter is dissolved, when the residue must
be washed and treated as the ordinary Diatomaceæ hereinafter
described.
Many diatoms are seen best when mounted in a dry state, the
minute markings becoming much more indistinct if immersed in
liquid or balsam; and for this reason those which are used as test
objects are usually mounted dry. Many kinds also are now prepared
in this way, as opaque objects to be examined with the lieberkuhn,
and are exquisitely beautiful. Others, however, are almost invariably
mounted in balsam; but as these will be again referred to in Chapter
III., and require the same treatment to fit them for the slide, it will
not be out of place to describe the cleaning and preparation of them
here. As before stated, there is much matter surrounding them
which must be got rid of before the “siliceous covering” can be
shown perfectly. As, however, we may first wish to become
acquainted in some degree with what we have to do, it is well to
take a small piece of talc, and place a few of the diatoms upon it.
This may be held over the flame of the spirit-lamp until all the
surrounding matter is burnt away, and a tolerable idea may be thus
obtained as to the quality of our treasure.
In some cases it is well to use this burning operation alone in
mounting specimens of diatoms, when they may be placed in their
natural state upon the thin glass, burnt for awhile upon the platinum
plate, hereafter described, and mounted dry or in balsam.
In the preparation and cleaning of Diatomaceæ, there is little
satisfaction unless these operations have been successfully
performed, as a very small portion of foreign matter seriously
interferes with the object. The mode of preparing them varies even
amongst the most experienced. It will be found, therefore, most
satisfactory to examine the principal of these separately, although it
may be at the risk of some little repetition.
The method which is the most frequently made use of is the
following:—Place the “gathering” containing the Diatomaceæ in a
small glass or porcelain vessel, add strong nitric acid, and, by the aid
of Bunsen’s burner or spirit-lamp, boil for some minutes. From time
to time a drop of the mixture may be put upon a slide, and
examined under the microscope to see if all foreign matter be got rid
of. When the valves are clean, the vessel containing them must be
filled with water, and the whole left for an hour or two, so that all
the diatoms may settle perfectly. The liquid must then be poured off
carefully, or drained away by the aid of a syphon, so that none of
the diatoms are removed with it. Indeed, it is well to examine the
liquid drained off each time with the microscope, as the finer forms
are frequently lost in the washings. The vessel must then be refilled
with pure water, allowed to settle, and drained as before. This
washing must be repeated until a drop being placed upon a slide
and evaporated leaves no crystals. When it is desirable to preserve
the diatoms in this state before mounting (which process will be
described in another place), they may be placed in a small phial with
a little distilled water.
There are many cases in which the above method will not effect
a perfect cleansing, as certain substances with which the diatoms
are frequently mixed are not soluble in nitric acid. For this reason the
following method is resorted to:—Take a quantity of the matter
containing the Diatomaceæ and wash first with pure water, to get rid
of all the impurities possible. Allow this to settle perfectly and decant
the water. Add hydrochloric acid gradually, and when all
effervescence has subsided, boil for some minutes by aid of the
lamp. When cool and the particles have subsided, decant the
hydrochloric and add nitric acid. The boiling must then be repeated
until a drop of the liquid when placed under the microscope shows
the valves or “frustules” clean. After allowing the diatoms to settle,
the acid must be decanted, and pure water substituted. The washing
must be repeated as in the former process until all the remains of
crystals or acid are removed, when the specimens may be preserved
in small phials as stated above.
Such are the usual modes of treating the Diatomaceæ, but there
are certain cases in which particular methods are required to give
anything like perfect results. Persons of great experience combine a
variety of treatments, and thus obtain better and more uniform
specimens. Perhaps it will be advantageous to give the young
student the process adopted by one of the most successful preparers
of these objects; but I will first state the different methods of
mounting the cleaned diatoms dry: how to employ Canada balsam
and fluid in their preservation will be elsewhere described.
It was before stated that the diatoms when cleansed might be
preserved in small phials of distilled water. When required for
mounting, shake the phial, and with a thin glass tube or rod take up
a drop of the fluid and spread it upon the surface of the slide in the
desired position. This must then be allowed to dry gradually, or by
the aid of the lamp if necessary, without being shaken or interfered
with, otherwise uniformity of dispersion will be prevented. When
thoroughly dry, a thin ring of one of the adhesive varnishes—gold-
size will be found as good as any—may be drawn round the diatoms,
and allowed to dry in a slight degree. The slide and thin glass cover
should then be warmed and the latter gently pressed upon the ring
of varnish until the adhesion all round is complete.
As some of the diatoms require object-glasses of extremely high
power, and, consequently, short focus, to show them, they must be
as close to the outer surface of the cover as possible. For this reason
they are sometimes placed upon the under side of the thin glass, as
follows. Clean the surfaces of the slide and cover, and with the rod
or pipe place the liquid containing the diatoms upon the thin glass,
and dry as before. Trace the ring to receive the cover upon the slide,
and when almost dry, warm both and proceed as above. Whichever
of these methods is employed, the outer ring of coloured varnish
may be applied as elsewhere described and the slide finished.
The diatoms are also sometimes mounted betwixt two thin
glasses, as described in Chapter I., so that the light by which they
are examined may receive as little interference as possible, and that
an achromatic condenser may be brought into focus under the slide.
Of the various modes of cleaning and mounting the
Diatomaceæ, I believe that the following may be safely
recommended as affording results of the very best quality. My friend,
Mr. T. G. Rylands, gave it to me as that which he prefers, and I can
safely say that his numerous slides are at least equal to any I have
ever seen. I will give it just as I received it from him, though there
may be some little repetition of what has been said elsewhere, as he
does not appropriate any part of it as his own. He says:—In this
branch of mounting, general rules alone can be laid down, because
the gatherings may contain iron, lime, fine silt, or vegetable matter
under conditions for special treatment, and consequently the first
step should be to experiment on various kinds.
In gathering diatoms much labour is saved by judgment and
care; hence it is desirable to get acquainted with them in their
growing condition, so that when recognised upon the sands or other
spots they may be carefully removed by the aid of the spoon or
small tin scoop before described. When growing upon algæ or other
plants, the plants and diatoms together may be carried home, in
which case they must be simply drained and not washed or pressed,
in order that the diatoms be not lost. As it is always desirable to
examine the gathering on the ground, a “Gairdner’s hand
microscope” with powers from 80 to 200 diameters will be found
very useful. The best gatherings are those which represent one
species abundantly. Those which are mixed may be rejected, unless
they are seen to contain something valuable or important, as the
object should be not so much to supply microscopical curiosities as
to collect material which is available for the study of nature.
The gathering when carried home should always be carefully
examined before anything is done with it; not only on account of the
additional information thus acquired, but also because it often
happens that a specimen should be mounted in fluid (see Chapter
IV.) in the condition in which it is gathered, as well as cleaned and
mounted in balsam (Chapter III.) and dry.
Where the gathering is taken from sand, the whole may be
shaken up in water as a preliminary operation, when much of the
sand will be separated by its own weight. The lime test, however,
should be applied, viz.—a small portion of hydrochloric acid, and if
there be effervescence it must be dissolved out by this means. From
Algæ and other weeds diatoms may be detached by agitating the
whole together in a weak solution of nitric acid—about one of pure
acid to twenty or thirty of water, as it must be sufficiently weak to
free the diatoms without destroying the matter to which they
adhere. The diatoms may then be separated by sifting through
coarse muslin, which will retain the Algæ, &c. The process of
cleaning will vary according to circumstances. Some gatherings
require to be boiled only a few minutes in nitric acid; but the more
general plan where they are mixed with organic or other foreign
matter, is to boil them in pure sulphuric acid until they cease to grow
darker in colour (usually from a half to one minute), and then to
add, drop by drop to avoid explosions, a cold saturated solution of
chlorate of potash until the colour is discharged, or, in case the
colour does not disappear, the quantity of the solution used is at
least equal to that of the acid. This operation is best performed in a
B
wide-mouthed ordinary beaker glass, a test-tube being too narrow.
The mixture whilst boiling should be poured into thirty times its bulk
of cold water, and the whole allowed to subside. The fluid must then
be carefully decanted and the vessel re-supplied once or twice with
pure water, so as to get rid of all the acid. The gathering may then
be transferred to a small boiling glass or test-tube, and—the water
being carefully decanted—boiled in the smallest available quantity of
nitric acid, and washed as before. This last process has been found
necessary from the frequent appearance of minute crystals, which
cannot otherwise be readily disposed of without the loss of a
considerable proportion of diatoms.

B
These glasses are round, about six inches
high, and usually contain about eight ounces.
They are rather wider at the bottom, tapering
gradually to the top, and may be generally
procured at the chemists, &c.
 I may here mention that the washing glasses used by Mr.
Rylands are stoppered conical bottles varying in capacity from two
ounces to one quart; the conical form being employed to prevent the
adherence of anything to the side; they are “stoppered” to render
them available in the shaking process about to be described.
The gathering, freed from acid, is now put into two inches depth
of water, shaken vigorously for a minute or two, and allowed to
subside for half an hour, after which the turbid fluid must be
carefully decanted. This operation must be repeated until all the
matter is removed which will not settle in half an hour. The fluid
removed should be examined by a drop being put upon a slide, as in
some cases very light diatoms have been found to come off almost
pure in one or more of these earlier washings. The quantity of water
and time of subsidence given may be taken generally, but may
require to be modified according to circumstances and the judgment
of the operator. By the repetition and variation of this process—the
shaking being the most important part—the gathering, if a pure one,
will be sufficiently clean. If, however, it contains a variety of species
and forms, it may require to be divided into different densities.
In some cases, however, it is best to divide the gathering as a
preliminary operation, which may be done by agitating it in a
quantity of water and decanting what does not readily subside. The
heavier and the lighter portions are then to be treated as two
separate boilings. But when the cleansing has been carried to the
above stage and this division is required, the plan must be
somewhat as follows:—The gathering must be shaken in a test-tube
with six inches of water, and then allowed to subside until one inch
at the top remains pure. About three inches are then to be carefully
withdrawn by a pipette, when the tube may be filled up and the
operation repeated. The three lower inches also may then be
decanted and examined. The gathering is thus divided into three
portions, viz.—that which was withdrawn by the pipette, that which
remained floating in the lower three inches of water in the tube, and
that which had settled at the bottom. An examination of these will
inform the operator how to obtain that particular density of
gathering which he desires, and how far it is worth while to refine
this process of elutriation; for in cases of necessity any one, or all
three, of these densities may be operated upon in the same way to
separate a particular diatom.
As occasional aids, it may be remarked, that in some cases
liquor ammoniæ may be used in place of water, as it often separates
fine dirt, which is not otherwise easily got rid of. Some fossil deposits
require to be treated with a boiling solution of carbonate of soda to
disintegrate them; but this operation requires great care, lest the
alkali should destroy the diatoms. Vegetable silicates also sometimes
require to be removed by a solution of carbonate of soda; but as the
frustules of the diatoms themselves are but vegetable silica, even
more care is required in this case. It may be well to mention, that
some diatoms are so imperfectly siliceous that they will not bear
boiling in acid at all. Some of these may be allowed to stand in cold
nitric acid some time, whilst others of a smaller and more delicate
character should, when possible, be treated with distilled water
alone.
We will now consider the mode of mounting the prepared
diatoms, which, if used dry (as described in this chapter), should be
carefully washed two or three times with the purest distilled water.
In this branch, as in every other, each collector gives preference to
that method in which he is an adept. Thus the diatoms may be
placed on the under side of the cover, to be as near to the object-
glass as possible, or upon the slide itself; and each plan has its
advocates. Whichsoever of these is used, nothing seems more
simple to the novice than a tolerably equal dispersion of the objects
upon the slide or cover; but this is by no means so readily
accomplished, consequently I give Mr. Rylands’ method, as his slides
are perfect in this respect also. He always places the diatoms upon
the thin glass cover. It is not sufficient, as is frequently thought, to
take a drop of liquid containing the cleansed material and spread it
upon the cover or slide, as without some additional precaution that
uniform and regular distribution of the specimens is not obtained
which is desirable. In order to effect this, let a drop of the cleansed
gathering be diluted sufficiently for the purpose—how much must be
determined in each case by experiment—and let the covers to be
mounted be cleaned and laid upon the brass plate. (See Chapter I.)
By means of a glass tube, about one-twelfth of an inch in diameter,
stopped by the wetted finger at the upper end, take up as much of
the diluted material as will form a moderately convex drop extending
over the whole cover. When all the covers required are thus
prepared, apply a lamp below the brass plate, and raise the
temperature to a point just short of boiling. By this means the covers
will be dried in a few minutes, and the specimens equally distributed
over the whole area. The spread of the fluid upon the covers is
facilitated by breathing upon them; and, to insure uniformity, care
must be taken to avoid shaking them whilst drying. The best plan is
to mount at least half a dozen at once.
Before mounting, Mr. Rylands always burns the diatoms upon
the glass at a dull red heat, whether they are used with balsam or
dry. This burning, he says, is not only an additional cleaning process,
but it effectually fixes the diatoms, and prevents them floating out if
mounted with balsam. The thinnest covers may be burnt without
damage if they are placed upon a small piece of platinum foil of the
size required, which should be about one-hundredth of an inch thick,
perfectly flat, and having three of its edges slightly bent over, so as
to prevent its warping with the heat. The small flame of a spirit-
lamp, or, where there is gas, a Bunsen’s burner, may be employed.
The cover should be shaded from direct daylight, that the action of
the flame may be observed more perfectly. Care must then be taken
to raise the temperature only to the dull red heat before mentioned.
The cover will then be in a fit state for mounting as required.
It has been stated in another place that it is assumed the
operator is not mounting diatoms simply as microscopic objects, but
as instructive specimens. It is not, therefore, sufficient to take a
single slide as all that is required, but to have the same diatom
prepared in as many ways as possible. The following are the
principal:—
 1. Mounted crude in fluid (see Chapter IV.).
2. Burnt crude upon the cover, and mounted dry or in
balsam (as before mentioned).
3. Mounted dry or in balsam (see Chapter III.), after the
cleansing process already described.

I will here give Mr. Rylands’ method of mounting them dry, the
fluid and balsam preparations being noticed in their respective
chapters. The slide with the ring of asphalt, or black varnish, should
have been prepared some weeks previously, in order to allow it to
dry thoroughly. When required, it must be held over the spirit-lamp
or Bunsen’s burner until the ring of varnish is softened. The burnt
cover, having been heated at the same time, must then be taken in
the forceps and pressed upon the softened varnish until it adheres
all round. When cold, an outer ring of asphalt completes the slide.
Such is the method which my friend Mr. T. G. Rylands employs in
the preparation of diatoms for the microscope. I have said enough
concerning his results. It is to be feared, however, that to some
these several modes of operation may appear lengthy and
complicated; but if read carefully, and the experiments tried, they
will be found simple enough in practice, and to occupy much less
time than an intelligible description would lead the novice to believe
necessary.
One of the most fertile as well as the most curious magazines of
Diatomaceæ is guano. The siliceous forms contained therein have
been devoured by sea-birds and passed through the stomach
uninjured, and after lying for ages may be cleaned and classified.
Many of these are not elsewhere met with, so that the student who
is desirous to enter into the study of Diatomaceæ must be instructed
as to the best mode of obtaining them from this source. The
particulars to be observed so closely resemble those before
mentioned in the treatment of the ordinary diatoms, that it will be
sufficiently explicit to give the outlines of the process. The guano
must be first washed in pure water, allowed to subside perfectly, and
the liquid then poured off. This must be repeated until the top fluid
is clear, and care taken not to decant the liquid until perfect
subsidence has taken place. The deposit must then be treated with
hydrochloric acid with a gentle heat for an hour or two, adding a
little fresh acid at intervals as long as it excites any effervescence
After this nitric acid must be substituted for the hydrochloric, and
the heat kept up to almost boiling-point for another hour at least,
adding a little fresh acid as before. When this ceases to act, the
deposit must be allowed to settle perfectly and the acid poured off.
All traces of the acid must now be washed away with pure water,
when the remains will be Diatomaceæ, the sand contained in the
guano, and a few other forms. Some of these may be mounted dry,
as before mentioned, but the greater portion should be put up in
Canada balsam as described in Chapter III.
Such is the ordinary method for the treatment of guano; but Mr.
Rylands’ mode of proceeding with ordinary Diatomaceæ (before
given) will be found equally successful with these deposits.
The fossil Infusoria (as they were formerly called) are now
termed Diatomaceæ, and are found in various parts of the world
—“Bermuda earth,” “Berg-mehl” from Norway, deposit from Mourne
Mountain in Ireland, &c. They are found in immense quantities, and
afford the microscopist innumerable objects. The same treatment as
that usually employed for the Diatomaceæ must be followed with
these deposits, but as they are sometimes obtained in hard masses,
disintegration is first necessary. To effect this, they are usually boiled
for a short time in diluted liquor potassæ, which will soon cause the
mass to fall into a mud-like deposit. Water must then be immediately
added, in order that all further action of the liquor potassæ may be
stopped, otherwise the objects searched for will be dissolved. For
this reason it is necessary to understand what substance is being
dealt with, because some deposits are much finer and acted upon
more readily than others.
In mounting these objects, some are so delicate that they are
almost invisible when balsam is used with them; they are therefore
usually mounted dry. Others, however, are much coarser, and may
be mounted in balsam like the Diatomaceæ mentioned in Chapter
III.
The common Infusoria cannot be mounted dry with any great
success, though a few may be placed upon the glass slide and
allowed to dry naturally, when their characters will be very well
shown. To obtain anything like a natural appearance, they must be
put up in fluid as in Chapter IV.
Next to the Diatomaceæ, no class of microscopic objects has
been more looked into of late than the Foraminifera. These animals
are almost all marine, having a jelly-like body enclosed in one or
more chambers of shell generally composed of carbonate of lime.
The shells are made with minute orifices, through which the
pseudopodia (false feet) are extended by which the animal is
enabled to lay hold of anything and draw itself along. From the
possession of these orifices they derive their name, as foramen
means a door or opening. They have been found in every depth of
sea hitherto sounded, each depth being abundant in certain species;
the lowest beds containing the greatest number of specimens,
though with less variation of kinds. In chalk they are found in a fossil
state, and may readily be shown (see Chapter III.); in limestone and
other hard stones they are abundant, and some mountains are
composed principally of these shells.
The methods of obtaining Foraminifera are various. Many may
be found upon seaweeds, which should always be examined as soon
as possible after gathering. They are found in masses upon some
coasts where the waves have carried and left them; but they are to
be found the most abundantly in sand or mud dredged from the
bottom of the sea. They must, however, be cleansed and separated
from the mass of impurity with which they are usually mixed. This
may be done in various ways, according to the nature of the
accompanying matter. If sand alone, as is frequently the case, the
whole mass must be thoroughly dried, and then stirred up in clean
water. The sand will soon subside by its own weight, but the
chambers of the Foraminifera, being filled with air, will float upon the
surface, and may be skimmed off. There is, however, one objection
to this mode of proceeding—some of these objects are so minute,
the chambers containing comparatively so small a quantity of air,
that they sink and are cast away with the refuse sand. On this
account it is preferable to take the trouble of searching certain
soundings under the microscope, using the camel-hair pencil, or
some other contrivance before mentioned, to extract those objects
which are required. To clean the Foraminifera, Professor Williamson
advises to transfer the specimens to an evaporating dish containing
a weak solution of caustic potash. This must be boiled for “some
moments,” when the organic matter will be entirely dissolved, and
the calcareous shells left free from impurity. They must now be well
washed in water, so that all alkaline matter may be entirely got rid
of.
If the specimens are in mud, we must proceed in a different
way:—Stir up the whole mass in water, and allow it to stand until the
heavier portion has sunk to the bottom; the water may then be
poured off and examined to see if there are any objects contained in
it. This process must be repeated until the water comes off quite
clear, when (if the search is for Foraminifera only) the solution of
caustic potash may be used as before mentioned. However the
soundings, &c., are cleaned, it is necessary to assort them under the
microscope with the camel-hair pencil or other contrivance, as it is
impossible to obtain them fit for mounting without undergoing this
process.
The sea soundings taken by order of Government are drawn
from the bottom in a kind of apparatus ingeniously made for the
purpose, and the sand, mud, &c., are brought up in their original
state. Common soundings, however, are taken by lowering a heavy
piece of lead coated with tallow, which consequently brings up a
small portion of the matter from the bottom. Mr. George Mosley, the
late Secretary of the Manchester Microscopic Society, obtained
numbers of the “scrapings” from the sounding leads. To make any
use of these it is, of course, necessary to free them from all traces of
the tallow. Mr. Dancer places the sounding in a basin and pours
boiling water upon it, which causes the melted grease to rise to the
surface. When cold, this may be removed, and the water carefully
decanted. The operation may be repeated until no grease appears,
when the water may be withdrawn and liquor ammoniæ used, which
will form a soapy solution with any remaining grease. This must be
treated with hot water for the final washing. Care must be taken lest
the finer forms be carried away in decanting the washing liquid.
Should it be wished to make certain as to this point, each washing
should be examined under the microscope. In some cases the
process of Mr. Dancer will prove sufficient. Mr. Dale, however, gives a
method of accomplishing the same result, which is much more
readily completed; and as the results cannot be found fault with, I
will here give it in full:—It is now well known that one of the
products obtained from the naphtha of coal-tar is a volatile, oily
substance, termed benzole (or, by French chemists, benzine), whose
boiling-point, when pure, is about 180° Fahrenheit, which is a
perfect solvent for fatty substances. In a capsule, previously warmed
on a sand-bath, Mr. Dale mixes with the tallow soundings benzole,
whose boiling-point may be about 200°, until sufficiently diluted so
as to run freely, pressing the lumps with a glass rod until thoroughly
mingled; the solution and its contents are then poured into a paper
filter, placed in a glass funnel; the capsule is again washed with
benzole, until the whole of the gritty particles are removed into the
filter. A washing-bottle is then supplied with benzole, and the
contents of the filter washed to the bottom until that liquid passes
off pure, which may be tested by placing a drop from the point of
the funnel on a warm slip of glass or bright platinum, when, if pure,
the benzole will evaporate without residue or tarnish; if grease be
present, the washings must be continued until they are free from it.
After rinsing through weak acid, or alcohol, for final purification, the
calcareous forms will be ready for mounting.
The filter and its contents may be left to dry spontaneously,
when the latter can be examined by the microscope. Should time be
an object, rapid drying may be effected by any of the usual
methods; one of which, recommended by Mr. Dale, is to blow a
stream of hot air through a glass tube held in the flame of a
Bunsen’s burner. The lower the boiling-point of the benzole, the
more readily can the specimens be freed from it. A commoner
quality may be used, but it is more difficult to dry afterwards.
Pure benzole being costly, this may appear an expensive
process; but, with the exception of a trifling loss by evaporation, the
whole may be recovered by simple distillation. The mixture of tallow
and benzole being placed in a retort in a hot-water, a steam, or a
sand bath, the benzole will pass into the receiver, and the tallow or
other impurities will remain in the retort. When the whole of the
benzole has distilled over, which is ascertained by its ceasing to drop
from the condenser, the heat is withdrawn and the retort allowed to
cool before the addition of fresh material. Half a dozen to a dozen
filters, each with its specimen, can be in process at the same time;
and the distillation of the recovered benzole progresses as quickly as
the filtration, which was practically proved on the occasion named.
Great caution in the use of benzole is to be taken in the approach of
lights to the inflammable vapour.
After the Foraminifera and calcareous forms have been removed,
the residue may be treated with acids and levigation in the usual
manner, to obtain siliceous forms and discs, if there are any present;
but to facilitate their deposition, and to avoid the loss of any minute
atoms suspended in the washings, I would suggest the use of
filtration. The conical filter is unsuitable, as the particles would
spread over too great a surface of paper; but glass tubes open at
both ends (such as broken test-tubes) will be found to answer, the
broad end covered with filtering paper, and over that a slip of muslin
tied on with a thread to facilitate the passage of the water and
prevent the risk of breaking the paper. Suspend the tube over a
suitable vessel through a hole cut in thin wood or cardboard, pour in
the washings which can be thus filtered and then dried. The cloth
must be carefully removed, the paper cut round the edges of the
tube, and the diatoms on the paper disc may be removed by a
camel-hair pencil or otherwise, ready for mounting. Thus many
objects may be preserved which would be either washed away or
only be obtained by a more tedious process.
Such is Mr. Dale’s method of cleaning the soundings from the
tallow, and as it thoroughly accomplishes its end, and is alike
effective and not injurious to Foraminifera and diatoms, it may be
safely recommended. The weak solution of caustic potash before
advised for Foraminifera, must not be used where it is desired to
preserve the diatoms, as they would certainly be injured, or
destroyed altogether, if this agent were employed.
In fixing the Foraminifera upon the slide, no better plan can be
followed than the “dry cells” and gum recommended in the early
parts of this chapter. Owing to their thickness and composition, most
of them are opaque objects only; but they are exquisitely beautiful,
and require no particular care, except in allowing the cell, &c., to be
perfectly dry, when the cover is placed upon it, or the damp will
certainly become condensed upon the inner side, and the
examination seriously interfered with.
Many of the Foraminifera require cutting into sections if it is
wished to examine the internal structure, &c.,—“decalcifying” is also
desirable in some cases;—both of these processes will be found
described at length in the chapter on Sections and Dissection.
When more than one specimen of some particular shell is
obtained, it is better to place them upon the slide in different
positions, so as to show as much of the structure as possible. I will
conclude this subject by quoting a passage from T. Rymer Jones:
—“It is, therefore, by no means sufficient to treat these shells as
ordinary objects by simply laying them on a glass slide, so as to see
them only from one or two points of view; they must be carefully
examined in every direction, for such is the diversity of form that
nothing short of this will be at all satisfactory. For this purpose, they
should be attached to the point of a fine needle, so that they may be
turned in any direction, and examined by reflected light condensed
upon them by means of a lens or side reflector. In many of the thick-
shelled species it will be necessary to grind them down on a hone
[see Chapter V.] before the number and arrangement of the internal
chambers is discernible; and in order to investigate satisfactorily the
minutiæ of their structure, a variety of sections, made in various
ways, is indispensable.”
Plants afford an almost inexhaustible treasury for the
microscope, and many of them show their beauties best when
mounted dry. When any of these also are to be mounted, care must
be taken that they are thoroughly dry, otherwise the damp will
certainly arise in the cell, and injure the object; and it may be here
mentioned that long after a leaf has every appearance of dryness,
the interior is still damp, and no way can be recommended of
getting rid of this by any quicker process than that effected by
keeping them in a warm room, as many leaves, &c., are utterly
spoiled by using a hot iron or other contrivance. The safest way is to
press them gently betwixt blotting-paper, which may be removed
and dried at short intervals; and though this may appear a tedious
operation, it is a safe one.
On the surface of the leaves, hairs and scales of various and
very beautiful forms are found, most of which display their beauties
best when removed from the leaf, and used with the polarizer. These
will be noticed in another place; but a portion of the leaf should
always be prepared in its natural form, to show the arrangement of
the hair or scales upon it; which must almost invariably be mounted
dry when used for this purpose. Many of them require very delicate
handling. The epidermis, or, as it is by some termed, the cuticle, is
the outer skin which lies upon the surface of the leaves and other
parts of most plants. This is composed of cells closely connected,
often bearing the appearance of a rude network. In many plants, by
scraping up the surface of the leaf, a thin coating is detached, which
may be torn off by taking hold of it with the forceps. The piece may
then be washed and floated upon a glass slide, where, on drying, it
will be firmly fixed, and may usually be mounted dry. Amongst the
most beautiful and easily prepared of these may be mentioned the
petal of the geranium, the cells of which are well defined and
amongst the most interesting.
 Closely connected with the leaves are the ANTHERS and POLLEN, of
which a great number are beautiful and interesting subjects for the
microscopist.
The mallow tribe will furnish some exquisite objects, bearing the
appearance of masses of costly jewels. These are usually dried with
pressure, but the natural form may be more accurately preserved by
allowing them to dry as they are taken from the flower, with no
interference except thoroughly protecting them from all dust.
Sometimes the anther is divided, so that the cell required to receive
them may be of as little depth as possible. The common mallow is a
beautiful object, but I think the lavatera is a better, as it shows the
pollen chambers well, when dried unpressed. The pollen is often set
alone, and is well worth the trouble, as it then admits of more close
examination. Often it is convenient to have the anther and pollen as
seen in nature on one slide, and the pollen alone upon another. The
former should be taken from the flowers before their full
development is attained, as if overgrown they lose much of their
beauty. Some pollens are naturally so dark that it is necessary to
mount them in Canada balsam or fluid, as described in other places;
but they are better mounted dry when they are not too opaque.
Here, too, we may also mention the SEEDS of many plants as
most interesting, and some of them very beautiful, objects, requiring
for the greater part but a low power to show them. Most of these
are to be mounted dry, as opaque objects, in cells suited to them,
but some are best seen in balsam, and will be mentioned in Chapter
III.
The Corallines, many of which are found on almost every coast,
afford some very valuable objects for the microscope. They must be
well washed when first procured, to get rid of all the salts in the sea-
water, dried and mounted in cells deep enough to protect them from
all danger of pressure, as some of them are exceedingly fragile. The
white ivory appearance which some of them present is given to them
by an even covering of carbonate of lime; and should it be desired to
examine the structure of these more closely, it may be accomplished
by keeping them for some time in vinegar or dilute muriatic acid,
which will remove the lime and allow of the substance being sliced in
the same way as other Algæ. (“Micrographic Dictionary,” p. 183.)
The Scales of Insects.—The fine dust upon the wings of moths
and butterflies, which is so readily removed when handled carelessly,
is what is generally called scales. To these the wing owes the
magnificent colours which so often are seen upon it; every particle
being what may be termed a distinct flat feather. How these are
placed (somewhat like tiles upon a roof) may be easily seen in the
wing of any butterfly, a few being removed to aid the investigation.
The form of them is usually that of the “battledore” with which the
common game is played, but the handle or base of the scale is often
short, and the broad part varies in proportionate length and breadth
in different specimens. The markings upon these also vary, some
being mostly composed of lines running from the base to the apex,
others reminding us of network—bead-like spots only are seen in
some—indeed, almost endless changes are found amongst them.
These scales are not confined to butterflies and moths, nor indeed to
the wings of insects. The different gnats supply some most beautiful
specimens, not only from the wings, but also from the proboscis,
&c.; whilst from still more minute insects, as the podura, scales are
taken which were at one time esteemed as a most delicate test. The
gorgeous colours which the diamond beetles also show when under
the microscope are produced by light reflected from minute scales
with which the insects are covered.
In mounting these objects for the microscope it is well to have
the part of the insect from which the scales are usually taken as a
separate slide, so that the natural arrangement of them may be
seen. This is easily accomplished with the wings of butterflies, gnats,
&c.; as they require no extraordinary care. In mounting the scales
they may be placed upon slides, by passing the wings over the
surface, or by gently scraping the wing upon the slide, when they
must be covered with the thin glass. Of course, the extreme tenuity
of these objects does away with the necessity of any cell excepting
that formed by the gold-size or other cement used to attach the
cover. The scales of the podura should be placed upon the slide in a
somewhat different manner. This insect is without wings, and is no
longer than the common flea. It is often found amongst the sawdust
in wine-cellars, continually leaping about by the aid of its tail, which
is bent underneath its body. Dr. Carpenter says:—“Poduræ may be
obtained by sprinkling a little oatmeal on a piece of black paper near
their haunts; and after leaving it there for a few hours, removing it
carefully to a large glazed basin, so that, when they leap from the
paper (as they will when brought to the light), they may fall into the
basin, and may thus separate themselves from the meal. The best
way of obtaining their scales, is to confine several of them together
beneath a wine glass inverted upon a piece of fine smooth paper;
for the scales will become detached by their leaps against the glass,
and will fall upon the paper.” These scales are removed to the slide,
and mounted as those from the gnats, &c. When the podura has
been caught without the aid of the meal, it may be placed upon the
slide, under a test-tube, or by any other mode of confinement, and
thus save the trouble of transfer from the paper before mentioned.
Another method is to seize the insect by the leg with the forceps and
drag it across the slide, when a sufficient quantity of scales will
probably be left upon it.
These scales are usually mounted “dry;” but Hogg recommends
the use of Canada balsam (Chapter III.) as rendering their structure
more definite when illuminated with Wenham’s parabolic reflector.
Some advise other methods, which will be mentioned in Chapter IV.
As most insects when undissected are mounted in Canada balsam,
the different modes of treatment which they require will be stated in
another place.
In mounting blood of any kind to show the corpuscles, or, as
they are often called, globules, which are round or oval discs, it is
but necessary to cover the slide on the spot required with a coating
as thin as possible and allow it to dry before covering with the thin
glass. There is a slight contraction in the globules when dried, but
not enough to injure them for the microscope. The shape of these
varies in different classes of animals, but the size varies much more,
some being many times as large as others.—Some of the larvæ skins
are beautiful objects; but, like many sections of animal and other
fragile matter, are difficult to extend upon the slide. This difficulty is
easily overcome by floating the thin object in clear water, immersing
the slide and when the object is evenly spread gently lifting it. Allow
it then to dry by slightly raising one end of the slide to aid the
drainage, and cover with the thin glass as other objects. The tails
and fins of many small fish may be mounted in a similar manner, and
are well worth the trouble.
A few objects which are best shown by mounting dry may be
here mentioned as a slight guide to the beginner, though some of
them have been before noticed;—many of the Foraminifera as
elsewhere described. Some crystals are soluble in almost any fluid or
balsam, and should be mounted dry; a few, however, deliquesce or
effloresce, which renders them worthless as microscopic objects.
The wings of butterflies and gnats, as before noticed, afford
many specimens wherewith to supply the cabinet of the young
student. A great variety also may be found amongst the ferns;
indeed, these alone will afford the student occupation for a long
time. On the under-side of the leaves are the reservoirs for the
“spores,” which in many instances somewhat resemble green velvet,
and are arranged in stripes, round masses, and other forms. The
spores are usually covered with a thin skin termed the indusium,
which is curiously marked in some specimens, often very like pollen-
grains. The manner in which these spores with all their
accompaniments are arranged, their changes and developments
afford almost endless subjects for study; different ferns presenting
us with many variations in this respect totally invisible without the
aid of the microscope. The hymenophyllums (of which two only
belong to England) are particularly interesting, and the structure of
the leaves when dried makes them beautiful objects, often requiring
no balsam to aid their transparency. Portions of the fronds of ferns
should be mounted as opaque objects, after having been dried
between blotting paper, when they are not injured by pressure; but
care must be taken to gather them at the right time, as they do not
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