Takayama Assay
FORS 420
Introduction: Takayama Assay is a test first discovered in the 1912. The assay
works by testing a sample with a pyridine and glucose reagent. In the presence of
blood, the reagent (pyridine and glucose) reacts with the heme group in blood and
produces hemochromogen crystals. Takayama assay is used for the confirmation of
blood not species origin.
Materials: Four Frosted Tip Slides
Four Cover Slips
Positive Sample
Unknown Samples
Slide warmer
Pyridine/Glucose Reagent
Micropipette
Compound Microscope
Procedure:
Gather four slides with frosted tips and four cover slips. Label each slide with
sample name, date, and initials.
Takayama Assay
FORS 420
- Gather each sample and give a detail description of the sample. Provide the
measurement of each sample and measurement of the cut piece take from the
sample.
-Place each of the cut pieces on the proper pre-labeled slide and place the cover
slip on top. Once the cover slide has been placed on top of the cut sample, circle
where the sample was placed on the backside of the slide. This helps you easily
locate the sample when viewing under a microscope.
Takayama Assay
FORS 420
- In a well ventilated area pipette the reagent under the cover slip, allow the
reagent to saturate the sample.
-After each sample has received the reagent place each slide onto the slide heater
(60-70ºC) for 2-5 minutes. After heating is complete each slide can now be viewed
under the microscope.
Takayama Assay
FORS 420
- When viewing under the microscope use the 10x and the 40x objectives. There
should be salmon pink crystals for positive blood samples. Record the
magnification used and location of the crystals using the measuring guide on the
microscope. This helps to refer back to the exact location of the crystals if needed.