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Universidade Federal do Rio de Janeiro

9 ARTICLES PUBLISHED IN JoVE

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Biochemistry

Lipid Index Determination by Liquid Fluorescence Recovery in the Fungal Pathogen Ustilago Maydis
Lucero Romero-Aguilar *1,2, Mónica Montero-Lomeli *3, Juan Pablo Pardo 2, Guadalupe Guerra- Sánchez 1
1Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, 2Facultad de Medicina, Universidad Nacional Autónoma de México, 3Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro

Here, we describe a protocol to obtain the lipid droplet index (LD index) to study the dynamics of triacylglycerols in cells cultured in high-throughput experiments. The LD index assay is an easy and reliable method that uses BODIPY 493/503. This assay does not need dispendious lipid extraction or microscopy analysis.

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Immunology and Infection

Three-Dimensional Imaging of the Vertebral Lymphatic Vasculature and Drainage using iDISCO+ and Light Sheet Fluorescence Microscopy
Laurent Jacob *1, Jose Brito *1,2, Jean-Leon Thomas 1,3
1Université Pierre et Marie Curie Paris, Sorbonne Université, Institut du Cerveau et de la Moelle Epinière, 2Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, 3Department of Neurology, Yale University School of Medicine

A protocol is presented combining tissue clearing with light sheet fluorescence microscopy (LSFM) to obtain three-dimensional and cellular resolution images of the lymphatic vessels and lymph nodes (LNs) collecting the cerebrospinal fluid (CSF) and spinal epidural fluid.

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Neuroscience

Visualizing Shifts on Neuron-Glia Circuit with the Calcium Imaging Technique
Matheus H. Tempone 1, Hercules R. Freitas 2, Clarissa S. Schitine 3, Ricardo A. de Melo Reis 1
1Laboratory of Neurochemistry, Institute of Biophysics Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, 2Laboratory of Neuroenergetics and Inborn Errors of Metabolism, Institute of Medical Biochemistry Leopoldo de Meis, Universidade Federal do Rio de Janeiro, 3Laboratory of Neurochemistry and Cell Biology, Institute of Life Sciences, Universidade Federal da Bahia

Cell calcium imaging is a versatile methodology to study dynamic signaling of individual cells, on mixed populations in culture or even on awakened animals, based on the expression of calcium-permeable channels/receptors that gives unique functional signatures.

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Neuroscience

Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents
Rafael Lani-Louzada 1, Carla Andreia Abreu 1, Victor G. Araújo 1, Mariana S. Dias 1, Hilda Petrs-Silva 1, Rafael Linden 1
1Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro

The goal of this protocol is to characterize a novel model of glaucomatous neurodegeneration based on 360° thermic cauterization of limbal vascular plexus, inducing subacute ocular hypertension.

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Biology

Quantitative Analysis of Viscoelastic Properties of Red Blood Cells Using Optical Tweezers and Defocusing Microscopy
Lucas Barreto *1,2, Fran Gomez *1,2, Pedro S. Lourenço 1,2, Douglas G. Freitas 1,2, Juliana Soares 1,3, Clemilson Berto-Junior 4,5, Ubirajara Agero 6, Nathan B. Viana 1,2, Bruno Pontes 1,2,3,7
1Centro Nacional de Biologia Estrutural e Bioimagem - CENABIO, Universidade Federal do Rio de Janeiro, 2Instituto de Física, Programa de Pós-graduação Multidisciplinar em Física Aplicada, Universidade Federal do Rio de Janeiro, 3Instituto de Biofísica Carlos Chagas Filho, Programa de Pós-graduação em Ciências Biológicas Biofísica, Universidade Federal do Rio de Janeiro, 4Faculdade de Farmácia, Universidade Federal do Rio de Janeiro - Campus Macaé, 5Faculdade de Medicina, Programa de Pós-Graduação em Endocrinologia, Universidade Federal do Rio de Janeiro, 6Instituto de Ciências Exatas, Departamento de Física, Universidade Federal de Minas Gerais, 7Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro

Here, an integrated protocol based on optical tweezers and defocusing microscopy is described to measure the rheological properties of cells. This protocol has wide applicability in studying the viscoelastic properties of erythrocytes under variable physio-pathological conditions.

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Biochemistry

Assessing Mitochondrial Function in Sciatic Nerve by High-Resolution Respirometry
Marcos A. Formiga-Jr 1, Juliana Camacho-Pereira 1
1Instituto de Bioquímica Médica Leopoldo de Meis, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro

High-resolution respirometry coupled to fluorescence sensors determines mitochondrial oxygen consumption and reactive oxygen species (ROS) generation. The present protocol describes a technique to assess mitochondrial respiratory rates and ROS production in the permeabilized sciatic nerve.

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Biochemistry

Preparation of Hard Palm Seeds for Matrix-Assisted Laser Desorption/Ionization-Imaging Mass Spectrometry Analysis
Gabriel R. Martins *1,2, Felipe L. Brum *1,2,3, Davi Marconi Miranda Carvalho da Silva *1,2, Livia C. Barbosa 3, Ronaldo Mohana-Borges 3, Ayla Sant'Ana da Silva 1,2
1Laboratório de Biocatálise (LABIC), Instituto Nacional de Tecnologia, 2Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, 3Centro de Espectrometria de Massas de Biomoléculas, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro

This protocol aimed to describe detailed guidance on the preparation of hard seed sample sections with low water content for MALDI-IMS analysis, maintaining analytes' original distribution and abundance and providing high-quality signal and spatial resolution.

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Biology

Studying Orthodontic Tooth Movement in Mice
José Alcides Almeida de Arruda 1, João Pacheco Colares 2, Mariana de Souza Santos 3, Victor Zanetti Drumond 4, Talita Martins 5, Carolina Bosso André 2, Flávio Almeida Amaral 6, Ildeu Andrade Jr. 7, Tarcília Aparecida Silva 8, Soraia Macari 2
1Department of Oral Diagnosis and Pathology, School of Dentistry, Universidade Federal do Rio de Janeiro, 2Department of Restorative Dentistry, School of Dentistry, Universidade Federal de Minas Gerais, 3Department of Physiology and Biophysics, Biological Science Institute, Universidade Federal de Minas Gerais, 4Private Clinic, 5Department of Metallurgical and Materials Engineering, School of Engineering, Universidade Federal de Minas Gerais, 6Laboratory of Immunopharmacology, Department of Biochemistry and Immunology, Biological Science Institute, Universidade Federal de Minas Gerais, 7Department of Orthodontics, School of Dental Medicine, University of Pittsburgh, 8Department of Oral Surgery, Pathology and Clinical Dentistry, School of Dentistry, Universidade Federal de Minas Gerais

Here, we present a protocol for studying orthodontic tooth movement (OTM), serving as a suitable model for investigating the mechanisms of bone adaptation, root resorption, and the response of bone cells to mechanical stimuli. This comprehensive guide provides detailed information on the OTM model, micro-computed tomography acquisition, and subsequent analysis.

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Biology

Non-invasive Skeletal Muscle Quantification in Small Animals Using Micro-computed Tomography
Alexander Pereira-Rosa 1, Thamires S. Oliveira 1, Matheus S. Ferreira 1, Rodrigo J. Vianna-Barbosa 2, Tula C. Wilmart-Gonçalves 2, Tânia M. Ortiga 1, Flavia F. Bloise 1
1Laboratório de Endocrinologia Translacional, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, 2Small Animal Imaging Unit, National Center for Structural Biology and Bioimaging, Universidade Federal do Rio de Janeiro

This protocol uses micro-computed tomography to enable the cost-effective quantification of lean mass, including skeletal muscle and visceral tissue, adipose tissue, and skeletal tissue in small animals. It distinguishes between lean and adipose tissue, which offers significant advantages for biomedical research, particularly in translational research in small animals.

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