Thin Sectioning of Slice Preparations for Immunohistochemistry

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.