THE4thEXPERIMENTIn situ display of peroxidasein the cell
THE 4th EXPERIMENT In situ display of peroxidase in the cell
[Aims] To master the method of the marrowsmear. To know in situ display technique ofthe peroxidase in the cell
[Aims] ⚫To master the method of the marrow smear. ⚫To know in situ display technique of the peroxidase in the cell
[principles]·The peroxidase can oxidate benzidineto be blue deposits or further brownandtheexistenceproducts,Sodistribution of the peroxidase in the cellare demonstrated indirectly
[ principles ] ⚫The peroxidase can oxidate benzidine to be blue deposits or further brown products, so the existence and distribution of the peroxidase in the cell are demonstrated indirectly
Contentsand Manipulations
[Contents and Manipulations]
Preparing smearExcide one end of the thighbone of the rabbitcut itopen lengthways, take out the marrow on the slide withbamboo stick and spread it (don't take too much ,asthin as possible), then dry it (by air-condition or airdry).Fixing2.Place it into 0.5% bluestone solution for 0.5min, thendry it.3.Reacting (Staining)Put it into the benzidine solution for 10mins, wash slideby the tap water(as gentle as possible) , then dry it.CounterstainingPut it into 1% Safranine solution for lmin, wash it bythe tap water, dry it , then examine sample undermicroscope(lowpowerhighpowandiooYoilimmersionlensesinturn
1. Preparing smear ⚫ Excide one end of the thighbone of the rabbit , cut it open lengthways, take out the marrow on the slide with bamboo stick and spread it (don’t take too much , as thin as possible) , then dry it (by air-condition or air dry). 2. Fixing ⚫ Place it into 0.5% bluestone solution for 0.5min, then dry it. 3. Reacting (Staining) ⚫ Put it into the benzidine solution for 10mins, wash slide by the tap water(as gentle as possible) , then dry it. 4. Counterstaining ⚫ Put it into 1% Safranine solution for 1min, wash it by the tap water，dry it , then examine sample under microscope.(low power, high power, and 100X oil immersion lenses in turn)
The arrows indicate the existence and distribution ofperoxidase in rabbit marrow cells with the blue orbrown granules ,× 1000
The arrows indicate the existence and distribution of peroxidase in rabbit marrow cells with the blue or brown granules， × 1000
Thereportofthe experiment The titleThe content?draw one marrow cell to show thedistributionoftheandexistenceperoxidase●Please turn in your reportbeforeleaving
⚫ The title ⚫ The content ⚫ draw one marrow cell to show the existence and distribution of the peroxidase ⚫ Please turn in your report before leaving. [The report of the experiment]
NOTE:3-4 students on duty are arranged by themonitor to clean the lab after experiment.Collect the garbage to the ash bin which is atthe corner nearby the toilet of this floor.·Clean the oil lens completely when you arethrough with the microscopeDonotdisassemble any parts of microscope·Please take back your microscope cards tothe monitor, then hand them over microscoperoom teacher when the experiment finished
NOTE: ⚫ 3-4 students on duty are arranged by the monitor to clean the lab after experiment. ⚫ Collect the garbage to the ash bin which is at the corner nearby the toilet of this floor. ⚫ Clean the oil lens completely when you are through with the microscope. ⚫ Do not disassemble any parts of microscope. ⚫ Please take back your microscope cards to the monitor, then hand them over microscope room teacher when the experiment finished