January is Cervical Cancer Awareness month, dedicated to raising awareness of cervical cancer. This includes the types of cervical cancer, what the symptoms are to look out for, cervical screening (smear testing), support groups available and information resources. It is important that there is an emphasis on how early detection can be the best prevention, especially when it comes to cervical cancer. Macmillan Cancer Support reports that “Each year, more than 3,200 women are diagnosed with cervical cancer in the UK.” A regular programme of Pap smear testing, with appropriate follow-up, can reduce cervical cancer incidences by up to 80%.
Rapid Pap Staining Kit
To support cytology gynaecology and non-gynaecology smears staining, we supply a Rapid Pap Staining Kit that enables complete sample processing in just a few minutes after sampling and produces excellent differentiation between cellular structures. Because of their simplicity of use and exceptional precision in diagnostics, reagents for staining according to Papanicolaou have substantially contributed to decreasing the occurrence of cervical cancer.
The staining procedure is simplified and conducted in a few minutes. It contains all the reagents necessary for sample processing – 95% alcohol as a fixative, deionized water, Hematoxylin HP, nuclear bluing reagent, OG-6 reagent and EA 50 reagent, as well as 100% alcohol for tissue dehydration and BioClear New (xylene substitute) for section clearing. It also contains a mounting medium of very low viscosity and optimal refractive index (BioMount New).
The reagents are placed in practical jars and sections may be directly immersed. They are placed in the order of use in the box, which lowers the possibility of contamination of reagents during staining and makes the kit quick and easy to use.
Below is a guide to how the kit can be used:
Preparing the cytological smear for staining
There are two methods of collecting and preparing the cytological samples:
- After collecting the cytological sample, place it on the microscope slide (VitroGnost), fixate it immediately with a fixative in a spray bottle (CitoSpray), dry it and keep until the staining process. Cytological sample may be fixated and kept until staining by immersing into 95% alcohol solution (Histanol 95) for a minimum of 30 minutes.
- Using liquid-based cytology method (LBC) and brush for collecting cytological samples, fixate the sample immediately (CitoFix, CitoFix in transport containers) by removing the brush head and immersing it in the fixative. At the beginning of processing the sample, isolate the cells from the fixative (one of the methods is to centrifuge the fixative) and place them on the microscope slide equally in a single layer. Cytological sample prepared in such a way is ready for staining.
Sample staining procedure
If the sample is dry and previously fixed using CitoSpray, it is necessary to keep it in a 95% alcohol solution (Histanol 95) for 10 minutes in order to remove polyglycols.
Start the staining process beginning from jar 1 containing Histanol 95. Continue with staining until the end (jar 19).
| Step | Jar | Method | Time/Process |
| 1 | 1 | Treat with 95% alcohol (Histanol 95) | 1 min |
| 2 | 2 | Treat with 95% alcohol (Histanol 95) | 10 dips |
| 3 | 3 | Rinse in deionized water | 10 dips |
| 4 | 4 | Stain using Hematoxylin HP, Pap 1A reagent | 30 sec |
| 5 | 5 | Rinse in deionized water | 10 dips |
| 6 | 6 | Nuclear bluing with Bluing reagent
Note: replace Bluing reagent with a fresh one (part of the kit) after each 50 sections |
1 min |
| 7 | 7 | Rinse in deionized water | 10 dips |
| 8 | 8 | Treat with 95% alcohol (Histanol 95) | 10 dips |
| 9 | 9 | Stain using OG-6, Pap 2B reagent | 2 minutes |
| 10 | 10 | Rinse using 95% alcohol (Histanol 95) | 10 dips |
| 11 | 11 | Rinse using 95% alcohol (Histanol 95) | 10 dips |
| 12 | 12 | Stain with EA50 reagent, Pap 3B | 4 minutes |
| 13 | 13 | Rinse using 95% alcohol (Histanol 95) | 10 dips |
| 14 | 14 | Rinse using 95% alcohol (Histanol 95) | 10 dips |
| 15 | 15 | Dehydrate using 100% alcohol (Histanol 100) | 10 dips |
| 16 | 16 | Dehydrate using 100% alcohol (Histanol 100) | 10 dips |
| 17 | 17 | Clear the section in BioClear New (xylene substitute) | 10 dips |
| 18 | 18 | Clear the section in BioClear New (xylene substitute) | 10 dips |
| 19 | 19 | Clear the section in BioClear New (xylene substitute) | 1 min |
Results
| Nuclei | Keratinized cells | Superficial squamous epithelial cell, erythrocytes, nucleoli, cilia |
Cytoplasm of all the other cell types (parabasal and intermediate squamous cells, columnar cells, polymorphonuclear leukocytes, lymphocytes, histiocytes, adenocarcinomas, undifferentiated carcinoma cells) |
|
blue-purple |
yellow-orange |
pink-red |
blue-green |
For more information about our Rapid Pap Staining Kit or to learn about our complete range of staining kits, speak to our team by calling +44 (0) 844 8080 900 or emailing labsupplies@solmedialtd.com.