MICROTOMES AND MICROTOMY

 

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MICROSCOPES AND MICROSCOPY

 

  

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DECALCIFICATION: PRINCIPLES AND PRACTICE

 

At the end of this lecture, students should be able to:    

1. Define decalcification and explain its prinicples

                                 2. Itemize and state common decalcifying agents with their merits and 

demerits

3. List and explain factors affecting decalcification

        4. Explain how to determine end point of decalcification

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Artefacts, Pigments And Minerals

 

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                                                         Objectives

                    At the end of this lecture, students should be able to:

                    1. Define artefacts, pigments and minerals

                    2. Differentiate endogenous from exogenous pigments

                    3. State and explain different methods of removing pigments

                    4. List methods of demonstrating pigments

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Wound Healing And Tissue Repair

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Objectives

At the end of this lecture, students should be able to:

1. Understand the basic concepts in cell and tissue regeneration

2. Identify the four phases of repair by connective tissues

3. Distinguish between healing by first intention and healing by second intention

4. List the three major roles of macrophages in wound healing

5. Differentiate between the morphology and histology of scar tissue

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INFLAMMATION AND TISSUE REPAIR -By Benard Solomon

 

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THEORY OF STAINING By Benard Solomon

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 https://docs.google.com/presentation/d/1DTlhjTeN3a2JgztJ82Kqx399V8ZHIy7Z-uJI1W9kH74/edit?usp=sharing

Tissues must be stained with dyes to make them visible.  The science behind staining is complex and multifaceted.  Different dyes stain different parts of tissues based on affinity and staining reactions.

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Haematoxylin Staining (H&E)

 

 

 Haematoxylin and Eosin Staining

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Use This To Solve Your Haematoxylin and Eosin staining problems

TISSUE PROCESSING

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Next: Staining. Watch Out.

11 Natural Dyes Used As Local Histological Stains

The concern for safety, cost-cutting, eco-friendliness and local availability is driving the quest to explore the abundant local plant resources in developing countries.  Researchers in these countries are now looking inwards to make use of resources in their domain to solve their problems. Recent works done in this regard are chronicled in this write up.

1.            Punica Granatum , (pomegranate): This was used for the staining of rat brain (Gharravi et al., 2006).

2.            Curcuma longa: The crude ethanolic extract and column chromatographic fraction of this local dye was used for the demonstration of collagen, muscle cells and red blood cells (Awvioro, 2007). It was also used for staining of seminiferous epithelium (Bassey et al., 2011).

3.            Kola nut (cola acuminata): This was used as eosin substitute to stain tissues (Shehu et al., 2012).

4.            Black plum fruit (Syzygium cumini): This was used as histological stain for rat hepatic tissue (Suabjakyong et al.,2011)

5.            Extract of butterfly pea flowers (Clitoria ternatea). This was used as histological stain for blood cells (Suebkhumpet and Sotthibandhu, 2012).

6.            Extract of dry leaves of henna (Lawsonia intermis).  This was used as histological stain for angiospermic stem (Jan et al. (2011).

7.            Red beet (Beta vulgaris) extract.  This was used as a histological stain for intestinal parasites (Al-Amura and co-researchers (2012).

8.            Black Mulberries.  This was used as histological stain for tissues (Tousson

 and AL-Behbehani (2011).

9.         Sorghum bicolor.  This was used as suitable counterstain for haematoxylin in histological sections (Omoowo, 2014). Also used for rat hippocampus cells demonstration  (Muhammed et al., 2016), liver and kidney (Benard et al., 2015) and brain cells (Benard et al., 2015).

10.       Baphia nitida (Camwood). This was used as histological stain for dentine tissues (Fasan, 

1977).

11.       Hibiscus sabdariffa.  This has been used as counter stain substitute to eosin ( Eman Hashim, 2006, Abdul-Afeez Ibnouf, 2014, Raheem et al., 2015).  It has also been explored as nuclear stain to substitute for haematoxylin (Benard, 2008, Benard et al., 2015, Benard et al., 2015, Benard et al., 2016, Muhammed et al., 2016, Agbede et al., 2017, Benard et al., 2017 and Omorodion and Achukwu, 2017).

These promising efforts should be followed up for the local laboratories to benefit from the translational research this findings offer.

References

1.         Abd-Alhafeez Ibnouf, Esam AbdulRaheem, Mohamed Seed Ahmed, and Dalia Dahab (2014). Assessment of staining quality of Roselle (Hibiscus Sabdariffa) on formalin-fixed paraffin-embedded renal tissue sections. Int J Cur Res Rev; 6 (21): 26-28.

2.         Agbede, M.B., Benard, S.A., Afolabi, O.O., Okoye, J.O., Bankole, J.K., Fowotade, A.A.,

Olutunde, O.A., Muhammed, O.A (2017):  The Use of Hibiscus Sabdariffa Extract as Nuclear Stain for Skin Morphology and Connective Tissue with Eosin Counterstain.  Sokoto Journal Of Medical Laboratory Science; 2(4):28-32

3.         Al-Amura, M. F.A., Hassen, Z.A. and ALMhanawi, B.H. (2012). Staining technique

for helminth parasites by use red beet (Beta Vulgaris L.) extract. Bas. J. Vet.

Res. 11(1): 283-292.

4.         Avwioro, O.G., Onwuka, S.K., Moody, J.O.,Agbedahunsi, J.M., Oduola, T., Ekpo, O.E. and Oladele, A.A. (2007). Curcuma longa extract as a histological dye for collagen fibres and red blood cells. J.Anat. 210: 600–603.

5.         Bassey R.B., Osinubi A.A., Oremosu A.A. (2012): Staining effect of Hibiscus sabdariffa extract on sperm cell morphology of Sprague –Dawley rats.  Journal of histotechonology 35(3): 110-113

6.         Benard S.A., Muhammed A.O., Fowotade A.A., Afolabi O.O., and Olutunde O.A.(2017).  Iron Roselle: A Progressive Nuclear Stain For Connective Tissue Of Skin. International Journal of Health Research and Innovation, vol.5. no.2, 25-31, ISSN: 2051-5057 (print version). Scienpress Ltd.

7.         Benard S.A., Muhammed A.O., Fowotade A.A., Afolabi O.O., Olutunde O.A. (2015): Sorghum  bicolor extract: A Suitable Counterstain In Hibiscus Extract Nuclear Staining of Liver And Kidney: African Journal Of Cellular Pathology 4: 13-16

8.         Benard S.A., Muhammed A.O., Fowotade A.A., Afolabi O.O., Olutunde O.A. (2015): Hibiscus  Sabdariffa Etract As Haematoxylin Substitute In The Histological Demonstration Of Brain Tissues: African Journal Of Cellular Pathology: 5:32-35

9.         Benard Solomon. Iron-Roselle (2008): A Progressive Nuclear Stain Substitute For Haematoxylin. J. Histotechnologyy. Vol. 31. 2, 31:57-59.

11.       Egbujo EC, Adisa OJ, and Yahaya AB. A Study of the Staining Effect of Roselle (Hibiscus Sabdariffa) on the Histologic Section of the Testis. Int. J. Morphol. 2008; 26(4):927-930.

12.       Eman A Hashim (2006). The use of watery extract of Kujarat flow­ers Hibiscus Sabdariffa as a natural histological stain. Iraqi J Med Sci. 5 (1): 29-33.

13.       Gharravi, A.M., Golarlipour, M.J., Ghorbani, R.and Khazaei, M. (2006). Natural dye for staining astrocytes and neurons. J.Neuro. Sci. (Turkisk) 23(3): 215-218.

14.       Ihuma JO, GH Asenge, JOK Abioye, and SK Dick (2012).  Applica­tion of Methanolic extracts from Hibiscus Sabdariffa Linn as a biological staining agent for some fungal species. In­ternational Journal of Plant, Animal and Environmental Sci­ences.; 2 (2): 254-9

15.       Okolie, N.J.C. 2008. Staining of ova of intestinal parasites with extracts of Hibiscus Sabdariffa and Azadirachta Indica. Inter. Sci. Res. J. 1(2): 116-119.

16.       Muhammed A.O., Olutunde O.A., Benard S.A., Muhammad Ahmad AT and Omoowo BT (2016):Hibiscus-Shorgum: A New Morphological Stain in Neuro-Histology: International Journal of Health Research and Innovation, vol.4. no.1, 31-38 ISSN: 2051-5057

17.       Omorodion N.T. and Achukwu P.U. (2017).  Investigation of Hibiscus Sabdariffa (Roselle) as Histological Stain and in Assessment of Bar Bodies. American Journal of Biomedical Sciences; 9(1), 15-19.

18.       Raheem M.A.,  Abd-Alhafeez O.I., Osman H.S., Hamza J.M (2015).  Using of Hibiscus Sabdariffa extract as a natural histological stain of the Skin. American Journal of Research Communication; 3(5), 211-216.

19.       Samanta, A. K., Agarwal, P. and Datta, S.(2007). Dyeing of jute and cotton fabrics

using jackfruit wood extract: Part-I :Effects of mordanting and dyeing process variables on colour yield and colour fastness properties. Indian J Fibre & Text Res. 32(12): 466-476.

20.       Suabjakyong, P., Romratanapun, S. and Thitipramote, N. (2011). Extraction of natural histological dye from black plum fruit (Syzygium cumini). J.Microsc. Soc. Thailand. 4(1): 13-15.

21.       Suebkhampet, A. and Sotthibandhu, P. (2012). Effect of using aqueous crude extract from butterfly pea flowers (Clitoria ternatea L.) as a dye on animal blood smear staining.

22.       Tousson, E. and Al-Behbehani, B. (2011).  Black Mulberries (Morus nigra) as a

natural dye for animal tissue staining. Animal Biol. 62(11): 49-56.

23.       Umar, A.A., Hena, S.A. and Wiam, I.M. (2012). Kola nut (Cola acuminata) extract as a substitute to histological tissue stain eosin. Sci. J. Vet. Adv. 1(2):33-37.