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A simple method for overcoming some problems when observing thick reflective biological samples with a confocal

C Rumio¹, M Morini, A Miani

¹University of Milan, Institute of Human Anatomy, Italy.

Journal of Microscopy

|January 1, 1995

Summary

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A novel device enhances confocal scanning laser microscopy for thick biological samples. This method reduces scanning depth and improves imaging of reflective or deep structures in tissues like skin and brain stem.

Area of Science:

Neuroscience
Histology
Microscopy

Background:

Confocal scanning laser microscopy (CSLM) faces challenges imaging thick, reflective biological samples.
Deep or opaque structures within specimens limit optical penetration and resolution.

Purpose of the Study:

To introduce a simple device and method to improve CSLM imaging of thick biological specimens.
To overcome limitations posed by sample reflectivity and depth in CSLM.

Main Methods:

A 'sandwich' style mounting technique using two coverslips for thick histological sections (human skin, rat brain stem).
Optical tomography performed from both sides by inverting the specimen.
Utilized standard Golgi-Cox and 'rapid Golgi' silver impregnation methods.

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Main Results:

The device effectively reduces the required scanning depth for CSLM.
Imaging of deep, opaque, or highly reflective structures within thick samples is significantly enhanced.
Successful visualization of intricate neural structures in Golgi-stained brain stem sections.

Conclusions:

The described 'sandwich' method with dual-sided optical tomography is a simple yet effective technique for CSLM of challenging biological samples.
This approach expands the utility of CSLM for detailed morphological studies of thick tissues.