k. Protocol guides and ethical issues
Guides and Protocols
For YouTube videos, you can also try related videos. Here we suggest just some, but try to find more!
Generals
- Biosecurity manual 1 and Biosecurity manual 2
- Code of good scientific practices (ICB/USP).
- Pipetting Handbook
- Pipetting guideline (only in Portuguese)
Animal care and handling
- 3-R Refinement (rat and mouse gavage, sample collection and intravenous administration) See this paper for complete information.
- Mouse Gavage Procedure (youtube video)
- Mouse Gavage Procedure 2 (youtube video)
- Rat Gavage Procedure (youtube video)
- Rat Gavage Procedure 2 (youtube video)
- Animal unit virtual tour (The University of Manchester) (video)
- Blood collection guide
- Blood sampling through saphenous vein (rat) (video)
- Immobilization of mouse for i.p. injection (video)
- Removal of visceral fats and adrenals in the Wistar rats (youtube video)
- Mouse Metabolic phenotyping Centers (Vanderbilt University) (youtube video)
- Perfusion of rat pancreas with collagenase for islet isolation (youtube video)
General information regarding methods
- Hemocytometer – Counting of cells (youtube video)
- Biosafety cabinet preparation to work and general safety instructions (youtube video)
- Cell Culture Basics (youtube video)
- Western Blot (youtube video)
- Western Blot (Thermo Fischer) recommendations (includes video)
- Bio-Rad Catalogue (see pages 176 for Protein Standards and page 183 for well’s volumes).
- Western Blot – “hall of shame“!
- Western Blot analysis with Image J (Part I) and (Part II) (Youtube video)
- Antibody validation (literature 1, literature 2 and literature 3).
- Quantification of blots using Image J. (youtube video)
- Analysis of data obtained with Image J (youtube video)
- ELISA (youtube video)
- Real-Time PCR (youtube video)
- Spectrophotometry Introduction (youtube video)
- Introduction to Flow Cytometry (manual) – Invitrogen video
- Fluorescence Spectrum Viewer A Multicolor Tool (BD biosciences tool)
- Isolation of mononuclear cells using Ficoll-Paque PLUS (manual) – Demonstration of the procedure (youtube video)
- Hyperinsulinemic-euglycemic Clamps in mouse (JOVE video)
- Link to download the lipoprotein calculator. Please, download it here.
Histology & Immunohistochemistry
- Removal of the rat pancreas for histomorphometry (YouTube video)
- Immunochemical protocols (Western, IHC, IF, ELISA) from ABCAM
- Fixation and tissue processing
- Fixation in formalin
- Complete immunohistochemistry guide
- Updated IHC application guide
- Microtome paraffin section I (youtube video)
- Microtome paraffin section II (youtube video)
- Frozen section in cryostat I (youtube video)
- Frozen section in cryostat II (youtube video)
- Histological techniques (Fiocruz)
- Histological techniques (UNICAMP)
- Histological panels (anato/histopathology).
- Examples: hepatic vacuoles (fat liver deposition); hepatic glucogenesis (glycogen deposition)
- ATLAS of Endocrine System Lesions in RATS.
- Determination of beta-cell mass using Image J.
- Electronic Microscope Atlas.
Animal ethics
- Brazilian guidelines for the use and care of animals for scientific and educational purposes – DBCA (updated in 2016)
- Brazilian guidelines (procedures with rodents) – CONCEA 2016
- Description of the invasive grades (G1, G2, G3 and G4).
- Brazilian guidelines for laboratory animal euthanasia – CONCEA (updated in 2018)
- For more regulations from CONCEA, access the site here
- Animal Research Advisory Committee Guidelines (Complete, from NIH). Here.
- Guideline for survival bleeding of Mice and Rats
- Guidelines for Endpoints
- Laboratory animals 1 and Laboratory animals 2
- Principles and Guidelines for Reporting Preclinical Studies
- Guide for the Care and Use of Laboratory Animals, 8th edition from the National Research Council
- For those who require the certification during undergrad. Please see the information here.
Good practices guidelines for researchers
