Histological Analysis of Lymph Nodes

For histological analysis, fixed lymph nodes were dehydrated in 100% ethanol and embedded in paraffin (Leica) using a Leica EG1150 tissue embedder. 7-μm-thick sections using a HM340E Thermo Scientific microtome were processed for hematoxylin-eosin (HE) (Leica) and immunohistochemistry staining as described (41 (link)). For immunohistochemistry of histology slides the following antibodies were used: anti-TER-119 (R&D Systems, MAB1125) and Alexa Fluor 488 conjugated goat anti-rat IgG antibody (Life Technologies A11006). Stained slides were mounted with Vectashield DAPI Mounting Medium (Vector Laboratories, H-1200). Tissue section samples were imaged using a Nikon Ni-U upright microscope (Nikon Instruments) using a 40x dry objective connected to a Nikon DS-Ri2 camera.

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Bálint L., Ocskay Z., Deák B.A., Aradi P, & Jakus Z. (2020). Lymph Flow Induces the Postnatal Formation of Mature and Functional Meningeal Lymphatic Vessels. Frontiers in Immunology, 10, 3043.

Publication 2019

EG1150 tissue embedder HM340E MAB1125 Alexa Fluor 488 conjugated goat anti-rat IgG antibody Vectashield DAPI Mounting Medium Nikon DS-Ri2

Alexa fluor 488 Anti igg Antibodies Antibody Dapi Dehydrated ethanol Embedded paraffin Eosin Goat Hematoxylin Immunohistochemistry Lymph nodes Microscope Microtome Tissue Vector

Corresponding Organization : Hungarian Academy of Sciences

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Variable analysis

independent variables

Dehydration in 100% ethanol
Embedding in paraffin (Leica) using a Leica EG1150 tissue embedder

dependent variables

Histological analysis of lymph nodes
Hematoxylin-eosin (HE) staining
Immunohistochemistry staining

control variables

7-μm-thick tissue sections using a HM340E Thermo Scientific microtome
Use of specific antibodies: anti-TER-119 (R&D Systems, MAB1125) and Alexa Fluor 488 conjugated goat anti-rat IgG antibody (Life Technologies A11006)
Mounting of stained slides with Vectashield DAPI Mounting Medium (Vector Laboratories, H-1200)
Imaging of tissue sections using a Nikon Ni-U upright microscope (Nikon Instruments) with a 40x dry objective connected to a Nikon DS-Ri2 camera

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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