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Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods

Introduction: Tear sampling is an attractive option for collecting biological samples in ophthalmology clinics, as it offers a non-invasive alternative to other invasive techniques. However, there are many tear sampling methods still in consideration. This study explores the suitability of Schirmer’...

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Main Authors:	Tham, May Ling, Mahmud, Aidalina, Abdullah, Maha, Md Saleh, Rafidah, Mohammad Razali, Amirah, Yoke, Kqueen Cheah, Mohd Taib, Niazlin, Kok, Lian Ho, Mahmud, Mazaya, Mohd Isa, Muhammad
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Published:	Springer Science and Business Media LLC 2023
Online Access:	http://psasir.upm.edu.my/id/eprint/110173/ https://www.cureus.com/articles/206096-tear-samples-for-protein-extraction-comparative-analysis-of-schirmers-test-strip-and-microcapillary-tube-methods
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spelling	my.upm.eprints.1101732024-09-04T04:20:31Z http://psasir.upm.edu.my/id/eprint/110173/ Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods Tham, May Ling Mahmud, Aidalina Abdullah, Maha Md Saleh, Rafidah Mohammad Razali, Amirah Yoke, Kqueen Cheah Mohd Taib, Niazlin Kok, Lian Ho Mahmud, Mazaya Mohd Isa, Muhammad Introduction: Tear sampling is an attractive option for collecting biological samples in ophthalmology clinics, as it offers a non-invasive alternative to other invasive techniques. However, there are many tear sampling methods still in consideration. This study explores the suitability of Schirmer’s test strip and microcapillary tube as reliable and satisfactory methods for tear sampling. Methods: Tear samples were collected from eight healthy volunteers using the standard Schirmer's test strip method with or without anesthesia and microcapillary tubes. The total tear protein concentrations were analyzed via spectrophotometry and bicinchoninic acid (BCA) protein assay. The protein profile was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal wetting length of Schirmer's strip and suitable buffer solutions were compared. Discomfort levels reported by participants and the ease of execution for ophthalmologists were also evaluated. Results: Tear samples exhibited typical protein profiles as shown by SDS-PAGE. The mean total protein obtained from an optimum wetting length of 20 mm using Schirmer's strip without anesthesia in phosphatebuffered saline (PBS) yielded substantial quantities of protein as measured by nanophotometer (220.20 ± 67.43 µg) and the BCA protein assay (210.34 ± 59.46 µg). This method collected a significantly higher quantity of protein compared to the microcapillary tube method (p=0.004) which was much more difficult to standardize. The clinician found it harder to utilize microcapillary tubes, while participants experienced higher insecurity and less discomfort with the microcapillary tube method. PBS used during the tear protein extraction process eluted higher tear protein concentration than ammonium bicarbonate, although the difference was not statistically significant. Using anaesthesia did not ease the sampling procedure substantially and protein quantity was maintained. Conclusion: Good quality and quantity of protein from tear samples were extracted with the optimized procedure. Schirmer's strip test in the absence of local anesthesia provided a standard, convenient, and noninvasive method for tear collection. Springer Science and Business Media LLC 2023 Article PeerReviewed Tham, May Ling and Mahmud, Aidalina and Abdullah, Maha and Md Saleh, Rafidah and Mohammad Razali, Amirah and Yoke, Kqueen Cheah and Mohd Taib, Niazlin and Kok, Lian Ho and Mahmud, Mazaya and Mohd Isa, Muhammad (2023) Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods. Cureus Journal Of Medical Science, 15 (12). art. no. e50972. pp. 1-12. ISSN 2168-8184 https://www.cureus.com/articles/206096-tear-samples-for-protein-extraction-comparative-analysis-of-schirmers-test-strip-and-microcapillary-tube-methods 10.7759/cureus.50972
institution	Universiti Putra Malaysia
building	UPM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Putra Malaysia
content_source	UPM Institutional Repository
url_provider	http://psasir.upm.edu.my/
description	Introduction: Tear sampling is an attractive option for collecting biological samples in ophthalmology clinics, as it offers a non-invasive alternative to other invasive techniques. However, there are many tear sampling methods still in consideration. This study explores the suitability of Schirmer’s test strip and microcapillary tube as reliable and satisfactory methods for tear sampling. Methods: Tear samples were collected from eight healthy volunteers using the standard Schirmer's test strip method with or without anesthesia and microcapillary tubes. The total tear protein concentrations were analyzed via spectrophotometry and bicinchoninic acid (BCA) protein assay. The protein profile was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal wetting length of Schirmer's strip and suitable buffer solutions were compared. Discomfort levels reported by participants and the ease of execution for ophthalmologists were also evaluated. Results: Tear samples exhibited typical protein profiles as shown by SDS-PAGE. The mean total protein obtained from an optimum wetting length of 20 mm using Schirmer's strip without anesthesia in phosphatebuffered saline (PBS) yielded substantial quantities of protein as measured by nanophotometer (220.20 ± 67.43 µg) and the BCA protein assay (210.34 ± 59.46 µg). This method collected a significantly higher quantity of protein compared to the microcapillary tube method (p=0.004) which was much more difficult to standardize. The clinician found it harder to utilize microcapillary tubes, while participants experienced higher insecurity and less discomfort with the microcapillary tube method. PBS used during the tear protein extraction process eluted higher tear protein concentration than ammonium bicarbonate, although the difference was not statistically significant. Using anaesthesia did not ease the sampling procedure substantially and protein quantity was maintained. Conclusion: Good quality and quantity of protein from tear samples were extracted with the optimized procedure. Schirmer's strip test in the absence of local anesthesia provided a standard, convenient, and noninvasive method for tear collection.
format	Article
author	Tham, May Ling Mahmud, Aidalina Abdullah, Maha Md Saleh, Rafidah Mohammad Razali, Amirah Yoke, Kqueen Cheah Mohd Taib, Niazlin Kok, Lian Ho Mahmud, Mazaya Mohd Isa, Muhammad
spellingShingle	Tham, May Ling Mahmud, Aidalina Abdullah, Maha Md Saleh, Rafidah Mohammad Razali, Amirah Yoke, Kqueen Cheah Mohd Taib, Niazlin Kok, Lian Ho Mahmud, Mazaya Mohd Isa, Muhammad Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods
author_facet	Tham, May Ling Mahmud, Aidalina Abdullah, Maha Md Saleh, Rafidah Mohammad Razali, Amirah Yoke, Kqueen Cheah Mohd Taib, Niazlin Kok, Lian Ho Mahmud, Mazaya Mohd Isa, Muhammad
author_sort	Tham, May Ling
title	Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods
title_short	Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods
title_full	Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods
title_fullStr	Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods
title_full_unstemmed	Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods
title_sort	tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods
publisher	Springer Science and Business Media LLC
publishDate	2023
url	http://psasir.upm.edu.my/id/eprint/110173/ https://www.cureus.com/articles/206096-tear-samples-for-protein-extraction-comparative-analysis-of-schirmers-test-strip-and-microcapillary-tube-methods
_version_	1811686055000145920
score	13.211869

Tear samples for protein extraction: comparative analysis of schirmer's test strip and microcapillary tube methods

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