Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> Shrimp
Histological techniques are essential for studying small crustaceans’, such as <i>Neocaridina</i> shrimp, anatomy and physiology. However, their small size and rapid tissue autolysis present challenges for fixation and processing. This study aimed to optimize histological methods for <...
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2025-06-01
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| author | Rafał Karol Wild Dobrochna Adamek-Urbańska Artur Witold Balicki Wiktoria Cieśla Jakub Przybyszewski Maciej Grzegorz Kamaszewski |
| author_facet | Rafał Karol Wild Dobrochna Adamek-Urbańska Artur Witold Balicki Wiktoria Cieśla Jakub Przybyszewski Maciej Grzegorz Kamaszewski |
| author_sort | Rafał Karol Wild |
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| description | Histological techniques are essential for studying small crustaceans’, such as <i>Neocaridina</i> shrimp, anatomy and physiology. However, their small size and rapid tissue autolysis present challenges for fixation and processing. This study aimed to optimize histological methods for <i>Neocaridina</i> shrimp by evaluating different protocols for fixation, decalcification, and enzymatic digestion. Shrimp were fixed using 10% neutral-buffered formalin (NBF) and Bouin’s or Davidson’s fluid with or without modifications such as trypsin digestion, decalcification, or abdomen removal. Tissue preservation, section quality, and staining properties were assessed. Davidson’s fluid consistently gave generally acceptable fixation results, with minimal autolysis and good tissue preservation. Trypsin digestion increased tissue damage and autolysis, particularly in the liver and pancreas. Decalcification improved the quality of the sections; however, it increased autolysis and resulted in less specific staining. The optimal protocol involved the removal of the abdomen, followed by fixation in Davidson’s fluid and decalcification, which resulted in rapid penetration of the fixative, minimal autolysis, and a beneficial effect on staining. This study highlights the importance of adapting histological methods to the specific characteristics of small crustaceans and provides a basis for future research on <i>Neocaridina</i> shrimp. Implementing these optimized techniques will improve the quality and reliability of histological analyses in crustacean research, deepening the understanding of their biology and facilitating their use as model organisms in various scientific fields. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-06-01 |
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| spelling | doaj-art-e0cbc045411940c294fd5b56185d43ec2025-08-20T03:26:16ZengMDPI AGAnimals2076-26152025-06-011512171510.3390/ani15121715Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> ShrimpRafał Karol Wild0Dobrochna Adamek-Urbańska1Artur Witold Balicki2Wiktoria Cieśla3Jakub Przybyszewski4Maciej Grzegorz Kamaszewski5Department of Animal Environment Biology, Institute of Animal Sciences, Warsaw University of Life Sciences (SGGW), 02-786 Warsaw, PolandDepartment of Animal Environment Biology, Institute of Animal Sciences, Warsaw University of Life Sciences (SGGW), 02-786 Warsaw, PolandDepartment of Animal Environment Biology, Institute of Animal Sciences, Warsaw University of Life Sciences (SGGW), 02-786 Warsaw, PolandDepartment of Animal Environment Biology, Institute of Animal Sciences, Warsaw University of Life Sciences (SGGW), 02-786 Warsaw, PolandDepartment of Animal Breeding, Institute of Animal Sciences, Warsaw University of Life Sciences (SGGW), 02-786 Warsaw, PolandDepartment of Animal Environment Biology, Institute of Animal Sciences, Warsaw University of Life Sciences (SGGW), 02-786 Warsaw, PolandHistological techniques are essential for studying small crustaceans’, such as <i>Neocaridina</i> shrimp, anatomy and physiology. However, their small size and rapid tissue autolysis present challenges for fixation and processing. This study aimed to optimize histological methods for <i>Neocaridina</i> shrimp by evaluating different protocols for fixation, decalcification, and enzymatic digestion. Shrimp were fixed using 10% neutral-buffered formalin (NBF) and Bouin’s or Davidson’s fluid with or without modifications such as trypsin digestion, decalcification, or abdomen removal. Tissue preservation, section quality, and staining properties were assessed. Davidson’s fluid consistently gave generally acceptable fixation results, with minimal autolysis and good tissue preservation. Trypsin digestion increased tissue damage and autolysis, particularly in the liver and pancreas. Decalcification improved the quality of the sections; however, it increased autolysis and resulted in less specific staining. The optimal protocol involved the removal of the abdomen, followed by fixation in Davidson’s fluid and decalcification, which resulted in rapid penetration of the fixative, minimal autolysis, and a beneficial effect on staining. This study highlights the importance of adapting histological methods to the specific characteristics of small crustaceans and provides a basis for future research on <i>Neocaridina</i> shrimp. Implementing these optimized techniques will improve the quality and reliability of histological analyses in crustacean research, deepening the understanding of their biology and facilitating their use as model organisms in various scientific fields.https://www.mdpi.com/2076-2615/15/12/1715crustacean histologyfixationdecalcificationprotocol optimization |
| spellingShingle | Rafał Karol Wild Dobrochna Adamek-Urbańska Artur Witold Balicki Wiktoria Cieśla Jakub Przybyszewski Maciej Grzegorz Kamaszewski Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> Shrimp Animals crustacean histology fixation decalcification protocol optimization |
| title | Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> Shrimp |
| title_full | Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> Shrimp |
| title_fullStr | Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> Shrimp |
| title_full_unstemmed | Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> Shrimp |
| title_short | Enhancing Histological Techniques for Small Crustaceans: Evaluation of Fixation, Decalcification, and Enzymatic Digestion in <i>Neocaridina</i> Shrimp |
| title_sort | enhancing histological techniques for small crustaceans evaluation of fixation decalcification and enzymatic digestion in i neocaridina i shrimp |
| topic | crustacean histology fixation decalcification protocol optimization |
| url | https://www.mdpi.com/2076-2615/15/12/1715 |
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