CRYOSTAT SECTIONING 1 TURN ON CRYOSTAT WITH “KEY” BUTTON—HOLD

104年04月共軛焦顯微鏡室製 冷凍組織細胞免疫螢光染色步驟 1 染色步驟 1 FIX CELLS OR CRYOSTAT
CRYOSTAT SECTIONING 1 TURN ON CRYOSTAT WITH “KEY” BUTTON—HOLD
MANUEL D’UTILISATION DU CRYOSTAT OXFORD SPECTROMAG PRÉPARATION

NCSX ENGINEERING DESIGN DOCUMENT DESIGN DESCRIPTION CRYOSTAT AND BASE
PANDA TS SPECTROMETER CRYOSTAT THICKNESS AND WEIGHT RENZO
SPECIFICATION OF THE AGATA THREE WAY CRYOSTAT GENERAL THE

Cryostat Sectioning

Cryostat Sectioning


  1. Turn on Cryostat with “Key” button—hold for a few seconds…then turn on light inside Cryostat.


  1. Adjust object temperature to -17 °C and cryostat temperature to -20 °C; if object is too cold, sections will wrinkle on stage (--According to Sadeq- start everyday at -20 °C/-20 °C for both OT and CT. Depending on the day you may need to adjust either OT or CT up or down --- usually one up and the other down).


  1. After mounting knife; place OCT sections in chamber and allow them ~ 20 minutes to equilibrate.


  1. Adjust cutting angle of knife – want optimally 5°; will scrape if too high, will give poor sections if too low; usually if set at 3° will actually cut at 5°.


  1. Align blade with clear, flip up stage, “anti-skid plate” if blade too high, tissue won’t go under plate; if blade too low, sections will wrinkle. When adjusting blade or plate, keep anti-skid plate up and out of the way of blade to avoid nicks in either.


  1. Remove specimen block and mount to circular stand using OCT. Place on holder until frozen.


  1. Label Gel-coated or Tespa Slides with pencil or marker—Shandon Lipshaw don’t fade with fixative.


  1. Trim sample to about 1 cm in length; adjust as necessary for optimal cutting.



  1. Cut slowly through block and when nice sections are obtained ations; if not continue to adjust; anti roll plate, angle of blade, temperature of cryostat and object…also may need to clean blade to obtain good sections.


  1. After obtaining 1 or 2 sections use a slight rocking motion to gently pick up sections with slide – start at bottom section and move in toward blade; --Don’t touch blade with slide.


  1. May subsequently freeze slides at -20 °C to -80 °C for Immunocytochemistry or In Situ Hybridization.





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