| E-Mail Address: | benardsolomona@yahoo.com |
Just recently, i completed my work on the staining of histological sections with extract of Hibiscus sabdarifa. The tissues were stained using (aq) solution of the extract at 56oC giving a silver-like staining appearance. The nuleus were stained with iron haematoxylin to give a black contrast.
This discovery could actually be expoited by the scientific community to determine the diagnostic value of this method in histopathology.
I want to welcome your comment and contribution on this issue.
send email to : benardsolomona@yahoo.com
DIFFERENTIAL STAINING OF HISTOLOGICAL SECTIONS WITH SOREL PLANT EXTRACT
ABSTRACT
This study describes the preparation and use of sorel plant extract for the differential staining of histological sections. The extract was prepared by heating several pieces of the sorel plant flower in water preceded by the addition of a mordant-potash alum. The resultant staining solution was used to stain sections obtained from formalin fixed tissues processed in an automatic tissue processor.
Sections from the gastro-intestinal tract, lymph node, uterus, liver and lung were stained.
Differential staining of the sections by the extract solution was obtained using different counterstains e.g. Van Gieson, Light Green and Eosin. Counterstaining with Van Gieson, and Light Green gave excellent results. Counterstaining with Methylene Blue gave a good result while Counterstaining with Eosin gave a poor result differentially.
It is suggested that sorel plant extract could be a new stain for histological diagnosis in Pathology Laboratories.
INTRODUCTION
Histological sections are distinctly and differentially stained when appropriate dys are employed to reveal structural details of tissues of interest. Such staining make possible the diagnosis of pathological changes or otherwise in various components of the tissues as a result of tissue interaction with the dye..
Dyes used could be natural of synthetic. Although most of the dyes employed in histopathology laboratories in this age are synthetic origin, yet, natural dyes still hold their fort where applicable e.g. Haematoxylin widely used for the staining of nucleus in tissues.
In Nigeria, dyes used for histopathological diagnosis are imported. Unfortunately, the unprecedented sled and devaluation of the major currency-Naira in the last decade had imparted negatively on the prices of medical analysis thus compromising their cost-effectiveness. It becomes imperative therefore for histotechnologists to look inward for alternative, local and cheaper resources for their analysis.
This paper describes the preparation and use of Sorel plant extract for the differential staining of histological sections.
MATERIALS NAD METHODS
Preparation of Sorel Plant Extract
The sorel plant is cultivated for the production of the ripe flowers which are found to have different species in most parts of West Africa. The extract used for this study was obtained from the red flowers of the plant.
Excess of 10g of Potassium alum was dissolved in 200ml of water by heating and shaking in a conical flask. The solution was brought to 60oC after which 10 pieces of the flowers were added to the solution which was further heated to boiling point rapidly. The solution was allowed to cool, filtered and kept in a clean, dry bottle ready for use.
Tissue Processing
10% formalin fixed tissues were processed automatically using the shandon tissue processor.
Processed tissues were embedded using tissue cassettes and rings. 5 microns of paraffin blocks were sectioned on a Leica Rotary Microtome.
Hydrated sections were stained overnight in already prepared extract solution, rinsed in running tap water and counterstained for 30 seconds in Van Gieson, Methylene Blue, Light Green and Eosin (1%) respectively. After staining, sections were dehydrated in alcohol, cleared in xylene and mounted in Canada Balsam.
RESULTS
Nuclear staining was achieved in all the sections stained. However, the degree of differentiation between the nucleus and cytoplasm were excellent with Van Gieson and Light Green counterstains-see table below:
DYE COMBINATION DEGREE OF DIFFERENTIATION COLOUR
Nucleus Cytoplasm
Extract + Light Green +++ Brown Green
Extract + Methylene Blue ++ Brown Blue
Extract + Van Gieson +++ Brown Pink
Extract + Eosin + obscured Red
Key
+++ Excellent
++ Good
+ Poor
DISCUSSION
The extract used for this study was obtained form the dry leaves of Rosel Calyx-a member of the hibiscus family. Its botanical name is Hibiscus Sabdariffa and it is found in most parts of West Africa (1). Histological sectios were not stained by the extract when applied to the tissues on its own. This finding corroborates the claim that natural dyes stain tissues poorly as they are easily washed out(2). This necessitates the use of an enhancement agent-mordant.
The principle of mordanting was to enhance the staining potential of the extract thus forming a dye-lake with affinity for nucleus. Potassium alum induced dye-lake bond always have nuclear affinity(2). This is particularly true for haematoxylin and the extract used for this study. It was also observed that the potassium alum-extract sequence resists acid decolorisation similar to celestine blue-haematoxylin sequence. The extract could therefore be a cheaper substitute for haematoxylin as a nuclear stain in techniques used for the demonstration of collagen fibres and other connective tissue fibres.
RERENCES
(1) J. Ayantuyi ( April 2, 2001), TELL Magazine pg 16
(2) D.J. Cook ( 1998) Cellular pathology, Butterworth Heiman, pg 63-95
Most Visited Biomedical Site 1
INTERNATIONAL FEDERATION OF BIOMEDICAL LABORATORY SCIENCE (WWW.ifbls.org)
This should be a most visit site for all biomedical practitioners all over the world for internet knowledge and education.
Home Page
The home page introduces the visitor to the site by giving the purpose of the site and the organisation running it. This page afford visitors opportunity of getting information about membership which could be accessed by clicking the red fonted link.
A pdf document on Biomedical laboratory science international is also conspicously placed for the perusal of interested visitors. ;you will miss alot of information if you ignore the link to this document.
Content
The educationn menu on the content column gives you international directories on medical laboratory education. The educational resources available are:
-Americal Society FOR Clinical Laboratory Science
-Americal Society FOR Clinical Pathology
-Canadian Society FOR Medical Laboratory Science
-Distance Education
-Med. Lab Continuing Education
-Quality systems
-Transfusion Medicine Resources
-Transfusion Quality Management Program
You can access these resources by a click of the mouse for example a click on the distance education option leads to 36 distance education sites in a jiffy. I particularly recomment the CSMLS link for intersted readers. you'll surely have more resources than you expect for self improvement and increased productivity.
LINKS
The links on this site are 21 in number but you may not have all the time in the world to borwse all the sites. a click on any of these links
-How to search the www
-Jobs
-Journals
will do you a lot of good.
To enjoy more go to www.ifbls.org
Most Visited Biomedical Site 2
AMERICAL SOCIETY FOR CLINICAL LABORATORY SCIENCE
This very colourful site is very rich in educaitonal materials as yo will find out in this write up.
HOME PAGE
e hoe page compacts all the resources you will most value. Apart from the content, ;you will see an invitational display for people seeking higher degree. You may click on this link if that is your priority now.
I recomment that you click on the 'current issues impacting on the profession and click
the 'bioterrorism preparedness link and ha a feel of what must be done to effectively control the consequences of theis phenomenon.
CONTENT
I do not want you to waste your time and money checking through the menu list. Go right away and click education. the page is rich in education materials and modules e.g. CD ROMS, Web Cournferences, Database info, Scholarships, education links and scientific assembly.
Colic on the educational links. On getting there, click MLS Distance Education prepared by web guru Pat Letendre.
The topics are;
-Genetics
-Immunology
-Methods
-ABO
-Rh
-Pretransfucsion testing
I recomment that you click on immunology to find out how it relates with Blood Transfusion.
LINKS
These are in the following categories
Regional Links
www.region2.vavalleyweb.com
www.utmem.edu
www.regioniv.ascls.org
www.tacls.org
State Societies
www.clsaonline.org
www.arscls.org
www.coscls.org
www.gscls.org
www.nj.ascls.org
MEDICAL LINKS
www.microbiol.org
www.medweb.emory.edu
www.healthtalk.com
INTERNATIONAL LINKS
www.aacc.org
www.att-info.org
www.asm.org
www.clma.org
To enjoy resources go to www.ascls.org
Last Updated on 2/26/08 by Benard Solomon.