Thin Sectioning of Slice Preparations for Immunohistochemistry
April 28th, 2007
The present method allows reproducible cryostat sectioning of small, difficult-to-manage, tissue pieces, such as biopsies and brain slices. We utilize a simple aluminum freezing stage to facilitate handling of tissue and a standard cryostat to routinely produce 5-10 micron serial sections from 400 micron thick brain slices.
Hi, my name is Jun Park Research fellow Harvard Medical School and also I am MD Student Y University College of Medicine Severance Hospital, EO Korea. These are materials for thin sectioning of slice preparations. I am preparing mold from tape four OCT platform.
I am feeling molded with OCT and freezing within cryostat or using crushed dry eyes am removing TAFE from around frozen OCT platform. I'm showing aligning marks on the freezing chalk and cryostat mounting stage and lock in chalk. I am sectioning through OCT platform.
Ontario surface is flat. I'm removing resurfaced OCT platform and placing on cryostat freezing stage. I'm placing this sample in OCT and this sample is previously cryo-preserved with 30%glycerol or sucrose in PBS.
This is the step for preparing freezing column with outer ring projecting about five millimeters above top of column forming. Well for OCTI Am carefully positioning tissue sample onto center of freezing column surface and slowly adding OCT until well is filled. I am surrounding freezing column with crushed dry eyes.
Tissue and OCT should completely freeze within 20 to 60 seconds. As preparation increases in temperature, the outer ring can be removed while the sample remains frozen. This is the step for sliding sample of freezing column sideways and placing in cryostat.
I am placing drop of OCT on surface of OCT platform and positioning spaceman applying form pressure spaceman will quickly freeze onto OCT platform. This is step of securing ch On to cryostat mounting stage with marks aligned. I am sectioning through OCT superficial to the tissue spaceman.
I am throwing mount thin sections onto glass slides and storing frozen or red room temperature in the reactions can be performed for tissue mounted on glass slides. Reagent is pulled onto slide can be gently agitated and may be covered if light sensitive subsequent stages of the reaction are easily performed by inverting. Slide into waste receptacle, weakening the slide, and then applying the next creation.
View the full transcript and gain access to thousands of scientific videos
Overview
This method enables reproducible cryostat sectioning of small, challenging tissue samples, including biopsies and brain slices. It employs a simple aluminum freezing stage for easier tissue handling and a standard cryostat to produce serial sections of 5-10 microns from 400 micron thick brain slices.
Key Study Components
Area of Science
- Neuroscience
- Histology
- Tissue Processing
Background
- Cryostat sectioning is essential for studying small tissue samples.
- Handling small biopsies and brain slices can be technically challenging.
- Standard methods may not yield reproducible results.
- Improved techniques can enhance the quality of tissue sections.
Purpose of Study
- To develop a reliable method for sectioning small tissue samples.
- To facilitate the handling of difficult-to-manage tissue pieces.
- To produce consistent and high-quality serial sections.
Methods Used
- Utilization of an aluminum freezing stage.
- Standard cryostat for sectioning.
- Production of 5-10 micron sections.
- Preparation of 400 micron thick brain slices.
Main Results
- Successful sectioning of small tissue samples.
- Reproducibility of section thickness.
- Improved handling of challenging tissue types.
- Consistent production of serial sections for analysis.
Conclusions
- The method enhances the ability to study small tissue samples.
- It provides a reliable approach for researchers in neuroscience.
- Future applications may include various types of tissue analysis.
