scholarly journals Immunocytochemical detection of lymphocyte surface antigens in fixed tissue sections.

1982 ◽  
Vol 30 (2) ◽  
pp. 171-174 ◽  
Author(s):  
M J Borowitz ◽  
B P Croker ◽  
J Burchette
Keyword(s):  
Surface Antigens ◽  
Fixed Tissue ◽  
Tissue Sections ◽  
Lymphocyte Surface ◽  
Immunocytochemical Detection

scholarly journals Tissue localization of lymphocyte surface antigens and receptors for immunoglobulin G Fc and complement in the chicken.

1982 ◽  
Vol 30 (3) ◽  
pp. 201-206 ◽  
Author(s):  
S Thunold ◽  
R Boyd ◽  
K Schauenstein ◽  
G Wick
Keyword(s):  
B Lymphocytes ◽  
B Lymphocyte ◽  
Surface Antigens ◽  
Mononuclear Phagocytes ◽  
Carbon Particles ◽  
Tissue Sections ◽  
Lymphocyte Surface ◽  
Rabbit Igg ◽  
Ig G

Frozen sections of chicken lymphoid organs were examined for lymphocyte surface antigens by antisera to T and B lymphocytes (ATS;ABS), and for the presence of immunoglobulin (Ig) G Fc and complement receptors (FcR;CR) by hemadsorption with sheep erythrocytes (E) coated with chicken IgG(EA), and E coated with rabbit IgG and chicken complement (EAC). In the spleen FcR positive cells were confined to the periellipsoidal sheaths and the germinal centers. CR positive cells were found in the same spleen areas, as well as in the medulla of bursal follicles. These lymphoid areas reacted strongly with ABS, but they also stained with neutral alpha-naphthyl butyrate esterase, and phagocytosed carbon particles were found in the periellipsoidal sheaths. Furthermore, in vivo treatment with cyclophosphamide, which resulted in pronounced B-lymphocyte depletion, did not affect FcR activity, but reduced CR activity significantly. These data indicate that the FcR activity demonstrated in tissue sections is mainly confined to mononuclear phagocytes, while the CR positive cells are mainly B lymphocytes.

Targeted ExSeq -- Tissue Preparation v2

2021 ◽  
Author(s):  
Anubhav Sinha ◽  
Yi Cui ◽  
Shahar Alon ◽  
Asmamaw T. Wassie ◽  
Fei Chen ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Human Tumor ◽  
Metastatic Breast ◽  
Tissue Type ◽  
Model Organisms ◽  
Tissue Preparation ◽  
Enzymatic Reactions ◽  
Fixed Tissue ◽  
Tissue Sections ◽  
Antibody Staining

This protocol accompanies Expansion Sequencing (ExSeq), and describes the tissue preparation for Targeted ExSeq. The steps described here are a generalization of the protocols used for figures 4-6 of the paper, and represent our recommendations for future users of the technology. Fig. 1 shows the structure of the protocol schematically. There are three possible tissue preparation routes described in this protocol that are applicable to different experimental systems. Option (A): harvesting tissue from model organisms that can be transcardially perfused with PFA, followed by sectioning using a vibratome. We typically use this workflow for work on mouse brain sections (see figures 4-5 of ExSeq paper). Option (B): transcardially perfusing with PFA, followed by cryoprotection and cryosectioning. We occasionally use this protocol for work on mouse brain sections. Option (C): snap-freezing fresh tissue (i.e., human tumor biopsy samples, or freshly harvested tissue from mice), followed by cryoprotection and cryosectioning (see figures 2 and 6 of ExSeq paper). The final result of options (A), (B), and (C) is the preparation of fixed tissue sections (either on a glass slide or free-floating). The protocols then briefly converge for optional antibody staining, treatment with LabelX, a chemical that enables anchoring of RNA to the expansion microscopy (ExM) hydrogel, followed by casting of the the ExM gel. There are minor differences in these steps between free-floating and slide-mounted tissue sections, which are noted in the individual steps. The next step, digestion, is tissue-type dependent and may require some optimization for your tissue type. We provide two potential options here: (1) a gentle digestion for tissues such as mouse brain, and (2) a harsh digestion for non-brain tissues such as tumor biopies. The protocols then converge again for the rest of the process. After digestion, the gels are expanded and re-embedded within a second non-expanding hydrogel to lock in the sample size. The carboxylates within the expansion gel are then chemically passivated, enabling enzymatic reactions to be performed within the gel. The samples are now ready for library preparation. In more detail: Steps 1-4 describe the preparation of reagents for downstream steps. The protocol begins either along options (A)/(B), the Transcardial PFA perfusion path (Step 5, continuing to vibratome sectioning in Steps 6-7 for option (A), or cryotome sectioning in Steps 9-10 for option (B)), or along option (C), the Fresh Frozen path (Step 8, continuing to cryotome sectioning in Steps 9-10). The protocols then converge for optional antibody staining (Step 11), followed by LabelX anchoring (Step 12), optional sample trimming (Step 13), and formation of the expansion microscopy gel (Step 14). The details of the digestion step are tissue-type dependent (Step 15). The protocol then concludes with expansion (Step 16), re-embedding (Step 17), passivation, and optional trimming (Steps 18-19). This protocol was used to profile human metastatic breast cancer biopsies as a part of the Human Tumor Atlas Pilot Project (HTAPP). The tissue for this work was collected (see HTAPP-specific tissue collection protocol). The tissue sections were then frozen, cryosectioned, post-fixed, and permeabilized (following steps 9-10). No antibody staining was performed (skipping optional step 11). The sections were then treated with LabelX and gelled (steps 12-14). The gels were then digested using the robust digestion option in steps 15-16. The samples were then re-embedded, passivated, and trimmed (following steps 17-19).

Enhanced DNA fragmentation in the thymus of spontaneously hypertensive rats

1999 ◽  
Vol 276 (6) ◽  
pp. H2135-H2140 ◽  
Author(s):  
Hidekazu Suzuki ◽  
Frank A. Delano ◽  
Neema Jamshidi ◽  
Dan Katz ◽  
Mikiji Mori ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Spontaneously Hypertensive Rat ◽  
Organ Injury ◽  
Fixed Tissue ◽  
Thymic Atrophy ◽  
Spontaneously Hypertensive ◽  
Tissue Sections ◽  
Dna Nicking ◽  
Wistar Kyoto

The mechanisms contributing to organ injury in hypertension have been incompletely defined. The thymus gland of the spontaneously hypertensive rat (SHR) shows significant atrophy at the age of 15 wk compared with its normotensive control, the Wistar-Kyoto rat (WKY). The aim of the present study was to examine the thymus of SHR for evidence of DNA nicking as one of the mechanisms for thymic atrophy. SHR and WKY were subjected to adrenalectomy or sham surgery at 12 wk and studied at 15 wk. Adrenalectomy served to normalize the blood pressure in the SHR. DNA nicking was detected by in situ nick-end labeling (ISEL) of fixed tissue sections. Tissue sections were treated with proteolysis, and terminal deoxyribonucleotidyl transferase was used to incorporate biotinylated deoxynucleotides into DNA nick end in situ. Separately, DNA fragmentation was evaluated by measuring the level of released mono- and oligonucleosomes to the cytoplasm. A higher number of thymic ISEL-positive cells and a higher level of cytoplasmic mono- and oligonucleosomes were observed in SHR than in WKY. After adrenalectomy the enhanced level of ISEL and cytoplasmic mono- and oligonucleosomes in SHR was reduced to the level in WKY. Dexamethasone treatment (0.05 mg ⋅ kg−1⋅ day−1) in WKY serves to decrease the thymus weight and significantly elevate the level of mono- and oligonucleosomes. Thus increased DNA fragmentation represents one of the mechanisms associated with thymic atrophy, a feature that reflects immune suppression in SHR.

In situ transcription: specific synthesis of complementary DNA in fixed tissue sections

Science ◽  
1988 ◽  
Vol 240 (4859) ◽  
pp. 1661-1664 ◽  
Author(s):  
L. Tecott ◽  
J. Barchas ◽  
J. Eberwine
Keyword(s):  
Fixed Tissue ◽  
Complementary Dna ◽  
Tissue Sections ◽  
Specific Synthesis

Lymphocyte Surface Antigens in Uveitis Patients

1999 ◽  
pp. 31-52
Author(s):  
R. Proen�a ◽  
A. Mota Pinto ◽  
M. Santos Rosa
Keyword(s):  
Surface Antigens ◽  
Lymphocyte Surface
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