Some microtechnical experiments and recommendations of new prestaining techniques

• Main Author: Wood, Edward D.
• Format: Others
• Published: Scholarly Commons 1947
• Subjects: Life Sciences; Zoology
• Online Access: https://scholarlycommons.pacific.edu/uop_etds/3532; https://scholarlycommons.pacific.edu/cgi/viewcontent.cgi?article=4528&context=uop_etds

The preparation and staining of biological structures for microscopic examination is of extreme importance to the science of biology since almost all phases of biological advance are, at least to some degree, dependent upon microtechnical methods. Nevertheless, research workers and students of biological science are sooner or later impressed with the inadequacy of many of the microtechnical methods used by them and their colleagues. In the field of microtechnique there is difficulty in finding standardized methods which will uniformly bring optimum results. Throughout most procedures there seem to be variables over which no definite controls have been established. Microtechniques have, in general, developed from trial and error efforts to obtain particular results in various biological researches where mastery of the techniques were of incidental or passing interest; and they are rarely based U!>On physio-chemical approach . One is impressed with the varied and often irrational methods recommended by different workers for presumably the same end results. As a consequence, the literature on the subject is scattered, fragmentary and unrelated to such an extent that a true science of microtechnique does not exist. From the writer's experience it was found that extensive perusal of microtechnical literature and use of procedures indicated therein, often did not result in the success anticipated. It was only after many selections and a great deal of trial and error that he was able to develop a dependable method for a particular purpose. The writer hopes that he has been able to find some of the underlying causes for the capricious reactions of tissues and that some of the haphazardness can be eliminated by the finding of some common denominators. Since there is a growing conviction that a large percentage of failures commonly attributed to staining techniques are actually due to pre-staining manipulations, it was decided to conduct some experiments on pre-staining techniques in an effort to lay a scientific foundation to further studies. The particular phenomena selected for specific treatment in this thesis were the physical alterations resulting from fixing fluids and from dehydration and dealcoholization reagents used prior to embedding, such as: (1) shrinkage, (2 ) hardening, (3 ) swelling , (4 ) crushing and other alterations of form.