Immuno electron microscopy ipre-embeddingjmethod
P@Cutting out of tissue

Q@The tissue is cut in detail.@TwTwQ


R@2.5%glutaraldehyde fixation@(2 hours or more in the temperature of 4 are fixed. )

S@It washes it with the phosphate buffer that adds the saccharose.

@Phosphate buffer that added 10% saccharoseiSAShores`overnightj
APhosphate buffer that added 20% saccharoseiSAShores`overnightj
BPhosphate buffer that added 30% saccharoseiSAShores`overnightj

T@The tissue is buried under the OCT compound, and it freezes with the acetone dry ice.

U@The frozen section of 6 microns is stuck on making silan coated glass.

V@The cut is dried.@iRoom temperature at RO minutesj

W@It washes it with cooled PBS.

X@@It washes it with 0.1% semicarbazide hydrochloric acid solution. @@@(It washes it once.)

PO@The aldehyde reaction is obstructed by using 0.1% semicarbazid hydrochloric acid solution. @(Room temperature for 1 hour)

PP@It washes it with the phosphate buffer solution.

PQ@It washes it by distilled water.

PR@It washes it with 200 time Immunosaver dilution solution.

PS@It puts on the constant temperature machine of soak 70 and the antigen retieval 16 hour processing is done to 200 time Immunosaver dilution solution.

PT@It washes it with the phosphate buffer solution.

PU@Immuno staining(LSABj

PV@It washes it with cooled PBS.

PW@It post-fixes by 1% osmium@acid. @S@Qhours

PX@It washes it with the phosphate buffer solution.

QO@Dehydration

QP@It embeded it in epoxy resin.

QQ@Ultra thin section @Electron microscope observation@@Electron micrograph