Question

Bone Assignment 1 - You received a small part from a bone lesion and the doctor needs the diagnosis very urgently. Describe how you will proceed with this specimen from start to end. 2- In a research laboratory you are doing an experiment on mice bone to find out enzymatic activity. What would be the best way to prepare your biopsy for this purpose. 3- you have decalcified a bone biopsy in a 5% HCl for classical H&E staining. Explain 3 steps that you should do to make sure that H & E staining will be of best possible qualities.

Answer 1

You received a small part from a bone lesion and the doctor needs the diagnosis very urgently. Describe how you will proceed with this specimen from start to end.

· The specimen will be collected in a container which is labeled with patient identifications such as Name, age, gender, medical record number, sample site etc. The specimen is then immersed in a formaldehyde will send to pathology lab for detailed examination and differential diagnosis (American Cancer Society, 2015).

· Pathologists then examine the sample without microscope which is called as gross examination. This is done to analyze the features of the specimens such as a size, color, measurement, appearance etc. (American Cancer Society, 2015).

· In case of sample specimen discussed in the above-mentioned scenario, the whole specimen will be looked under a microscope (American Cancer Society, 2015).

· Then the specimen will be put in cassettes, which is a small container in order to secure the specimen while processing (American Cancer Society, 2015).

· After processing which may take from hours to overnights the specimen will be placed in a mold with hot paraffin wax which solidify on cooling to protect the specimen (American Cancer Society, 2015).

· The embedded tissue in the paraffin block will be sliced and placed on glass slides which are later immersed into various stains and dye for further examination. The color change due to dyes makes it easier for the examination (American Cancer Society, 2015).

In a research laboratory you are doing an experiment on mice bone to find out enzymatic activity. What would be best way to prepare your biopsy for this purpose?

The best way to prepare mice bone biopsy for enzymatic activities is mentioned below.

· The tissue can be minced and homogenized into four volumes in Phosphate buffer.

· The homogenate can be sonicated by cooling for 3 minutes.

· Then the sonicated homogenate of the specimen can be centrifuged for half an hour (Biological Research, 2009)

You have decalcified a bone biopsy in a 5% HCL for classical H&E staining. Explain 3 sets that you should do to make sure that H & E staining will be of best possible qualities.

· Stain in 200ul hematoxylin solution and incubate at room temperature for 5 minutes

· Stain in 400ul Eosine solution for half a minute

· Dehydrate in absolute alcohol, two changes of two minutes each. Then clear in Xylene two changes in 10 minutes each.