Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia Morphology

In homeostasis, the glial cells support pivotal functions, such as neuronal differentiation, neuroprotection, nutrition, drug metabolism, and immune response in the central nervous system (CNS). Among these cells, astrocytes and microglia have been highlighted due to their role in the pathogenesis o...

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Main Authors: Alana Alves Farias, Ana Carla dos Santos Costa, Jéssica Teles Souza, Érica Novaes Soares, Cinthia Cristina de Oliveira Santos Costa, Ravena Pereira do Nascimento, Silvia Lima Costa, Victor Diogenes Amaral da Silva, Maria de Fátima Dias Costa
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Neuroglia
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Online Access:https://www.mdpi.com/2571-6980/6/2/16
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author Alana Alves Farias
Ana Carla dos Santos Costa
Jéssica Teles Souza
Érica Novaes Soares
Cinthia Cristina de Oliveira Santos Costa
Ravena Pereira do Nascimento
Silvia Lima Costa
Victor Diogenes Amaral da Silva
Maria de Fátima Dias Costa
author_facet Alana Alves Farias
Ana Carla dos Santos Costa
Jéssica Teles Souza
Érica Novaes Soares
Cinthia Cristina de Oliveira Santos Costa
Ravena Pereira do Nascimento
Silvia Lima Costa
Victor Diogenes Amaral da Silva
Maria de Fátima Dias Costa
author_sort Alana Alves Farias
collection DOAJ
description In homeostasis, the glial cells support pivotal functions, such as neuronal differentiation, neuroprotection, nutrition, drug metabolism, and immune response in the central nervous system (CNS). Among these cells, astrocytes and microglia have been highlighted due to their role in the pathogenesis of several diseases or due to their role in the defense against several insults (ex., chemicals, and pathogens). <i>In Vitro</i> cytological analysis of astrocytes and microglia has contributed to the understanding of the role of morphological changes in glial cells associated with a neuroprotective or neurotoxic phenotype. Currently, the main tools used for the investigation of glial cell morphology in culture are phase contrast microscopy or immunolabeling/fluorescence microscopy. However, generally, phase contrast microscopy does not generate images with high resolution and therefore does not contribute to visualizing a single cell morphology in confluent cell cultures. On the other hand, immunolabeling requires high-cost consumable antibodies, epifluorescence microscope or confocal microscope, and presents critical steps during the procedure. Therefore, identifying a fast, reproducible, low-cost alternative method that allows the evaluation of glial morphology is essential, especially for neuroscientists from low-income countries. This article aims to revise the use of Rosenfeld’s staining, as an alternative low-cost and easy-to-reproduce method to analyze astrocytic and microglial morphology in culture. Additionally, it shows Rosenfeld’s staining as a valuable tool to analyze changes in neural cell morphology in toxicological studies.
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spelling doaj-art-9dd4f735d9294008a5877b42e28796592025-08-20T03:29:45ZengMDPI AGNeuroglia2571-69802025-04-01621610.3390/neuroglia6020016Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia MorphologyAlana Alves Farias0Ana Carla dos Santos Costa1Jéssica Teles Souza2Érica Novaes Soares3Cinthia Cristina de Oliveira Santos Costa4Ravena Pereira do Nascimento5Silvia Lima Costa6Victor Diogenes Amaral da Silva7Maria de Fátima Dias Costa8Laboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilLaboratory of Neurochemistry and Cellular Biology, Institute of Health Sciences, Federal University of Bahia, Av. Reitor Miguel Calmon S/N, Salvador 40231-300, BrazilIn homeostasis, the glial cells support pivotal functions, such as neuronal differentiation, neuroprotection, nutrition, drug metabolism, and immune response in the central nervous system (CNS). Among these cells, astrocytes and microglia have been highlighted due to their role in the pathogenesis of several diseases or due to their role in the defense against several insults (ex., chemicals, and pathogens). <i>In Vitro</i> cytological analysis of astrocytes and microglia has contributed to the understanding of the role of morphological changes in glial cells associated with a neuroprotective or neurotoxic phenotype. Currently, the main tools used for the investigation of glial cell morphology in culture are phase contrast microscopy or immunolabeling/fluorescence microscopy. However, generally, phase contrast microscopy does not generate images with high resolution and therefore does not contribute to visualizing a single cell morphology in confluent cell cultures. On the other hand, immunolabeling requires high-cost consumable antibodies, epifluorescence microscope or confocal microscope, and presents critical steps during the procedure. Therefore, identifying a fast, reproducible, low-cost alternative method that allows the evaluation of glial morphology is essential, especially for neuroscientists from low-income countries. This article aims to revise the use of Rosenfeld’s staining, as an alternative low-cost and easy-to-reproduce method to analyze astrocytic and microglial morphology in culture. Additionally, it shows Rosenfeld’s staining as a valuable tool to analyze changes in neural cell morphology in toxicological studies.https://www.mdpi.com/2571-6980/6/2/16Rosenfeld’s stainingastrocytesmicrogliaPC-12 cellsmorphology
spellingShingle Alana Alves Farias
Ana Carla dos Santos Costa
Jéssica Teles Souza
Érica Novaes Soares
Cinthia Cristina de Oliveira Santos Costa
Ravena Pereira do Nascimento
Silvia Lima Costa
Victor Diogenes Amaral da Silva
Maria de Fátima Dias Costa
Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia Morphology
Neuroglia
Rosenfeld’s staining
astrocytes
microglia
PC-12 cells
morphology
title Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia Morphology
title_full Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia Morphology
title_fullStr Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia Morphology
title_full_unstemmed Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia Morphology
title_short Rosenfeld’s Staining: A Valuable Tool for <i>In Vitro</i> Assessment of Astrocyte and Microglia Morphology
title_sort rosenfeld s staining a valuable tool for i in vitro i assessment of astrocyte and microglia morphology
topic Rosenfeld’s staining
astrocytes
microglia
PC-12 cells
morphology
url https://www.mdpi.com/2571-6980/6/2/16
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