FHML course Immunology Virtual microscopy Lymphoid tissues ‘Activity centers in the immune response’ Dr. A.M. Duijvestijn

Part 2: Tonsils (1) Tasks and questions: -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. T1: What are the names of the 2 main types of tonsils in the oro-naso-pharynx? Answer:        Q. T2: What type of epithelium does the mucosa of this tonsil consists of? So, which type of tonsil do we see here? Q. T3: What is the name for the collection of different lymphoid tissues in the back of the oronasopharynx? Q.T4: What is the function of the crypts of the tonsil?

Part 2: Tonsils (2) Tasks and questions: -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. T5: Make a screenshot of the whole tonsil section and copy-paste it in the frame  .First click in the frame before pasting Mark the lumen of the nasopharynx with an asterisk.

Part 2: Tonsils (3) Tasks and questions: -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q.T6: Select and magnify in the section a tonsil surface area showing the ‘finger like’ lymphoid tissue and crypts. Select and copy-paste this area in the available frame of this power point (PPT) slide. First click in the frame before pasting Q.T7: Search for an area in the tonsil with a clear mucosal lympho-epithelial network. Magnify, select and copy-paste this to your PPT.

Part 2: Tonsils (4) Tasks and questions: -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q.T8: Select 2 B-cell follicles to add to your PPT. Indicate whether or not they contain a germinal centre. Q.T9: What is the function of the germinal centre ? Answer: Q.T10: Search for an ‘interfollicular’ (T-cell area) in which HEVs can be recognized. Select and magnify an HEV to add to your PPT. Q.T11: What is the function of an HEV? Answer: -> After answering the questions go back to the website and continue with part 2: Appendix

Part 3: Appendix (1) Tasks and questions: -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. A1: Make a screenshot of the whole appendix section and copy-paste it in the frame .

Part 3: Appendix (2) Tasks and questions: -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. A2: How many follicles with germinal centres does the preparation approximately contain? Answer: Q. A3: Which special cells in the epithelium at the surface of the lymphoid tissue can transport small antigen particles from the intestinal lumen to the underlying lymphoid structures? Q. A4: Which cells further process the transported antigen in the lymphoid tissue?

Part 3: Appendix (3) Tasks and questions: -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. A5: Select and magnify for your PPT an area of the appendix in which villi, crypts and underlying lymphoid tissue can be recognized. Indicate what is what. Q. A6: Select and magnify for your PPT 2 or more B-cell follicles with interfollicular T-cell areas. Indicate the presence of a germinal centre with an asterisk. Q. A7: Magnify a representative HEV out of the T-cell area and paste it in your PPT -> After answering the questions go back to the website and continue with part 3: Peyer’s patches

Part 4: Peyer’s patches (1) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. P1: Make a screenshot of the whole section and copy-paste it in the frame .

Part 4: Peyer’s patches (2) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. P2:  In which part of the small intestine do we find the Peyer's patches? Answer: Q. P3:  Which 3 processes relevant for the humoral (antibody) immune response take place in the germinal centres? Q. P4: Select and magnify the area of the section that contains the Peyer’s patch. Copy-paste it to your PPT. Q. P5: How many B-cell follicles does this PP contain, and in how many of them do you recognize a germinal centre ? Answer: Q. P6: Point out in the figure the germinal centre(s) in the B-cell follicle(s), and the T-cell area(s). -> After answering the questions go back to the website and continue with part 5: Lymph nodes; General histology

Part 5: Lymph nodes; General histology (1) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. L1: Make a screenshot of the whole lymph node section and copy-paste it in the frame .

Part 5: Lymph nodes; General histology (2) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. L2: Where do the afferent (supplying) lymph vessels enter the lymph node? Answer: Q. L3: Select and magnify an outercortical area with a primary B-cell follicle, and copy-paste it to your PPT. Q. L4: Do the same for a secondary follicle, point out the germinal centre with an asterisk.

Part 5: Lymph nodes; General histology (3) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. L5: Select and magnify a paracortical area in which 1 or more HEV can be seen. Copy-paste it to your PPT, and indicate the HEV(s). Q. L6: Select and magnify a medulla area at the hilus side. Copy/paste it to your PPT, and indicate with an arrow the large vessels at the hilus. Q. L7: Which 3 large vessels can be found at the hilus? Answer: -> After answering the questions go back to the website and continue with part 6: Lymph nodes; Immunohistology

Part 6A: Lymph nodes; Immunohistology ‘Reticulum staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh1: What are the circular structures in the parenchyma of the node ? Answer: Q. LIh2: Select, magnify, and copy-paste a capsular area with clear subcapsular sinus to your PPT.  Q. LIh3: Select, magnify high and copy-paste an area in the parenchyma in which the fine structure of the reticular network can be distinguished. -> After answering the questions go back to the website and continue with part 6B: Lymph nodes; Immunohistology ‘Macrophage staining’

‘Macrophage staining’ Part 6B: Lymph nodes; Immunohistology ‘Macrophage staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh4: What is the distribution of scavenger macrophages in the lymph node ? Answer: Q. LIh5: In which area(s) of this lymph node do you find the highest number of these macrophages?   Q. LIh6: What is the main function of these macrophages in the lymph node? Answer: Q. LIh7: Select and magnify the area with the highest number of macrophages and copy-paste it to your PPT. -> After answering the questions go back to the website and continue with part 6C: Lymph nodes; Immunohistology ‘B-cell (IgM) staining’

‘B-cell (IgM) staining’ Part 6C: Lymph nodes; Immunohistology ‘B-cell (IgM) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh8: In which areas other than the outer cortex are B-cells present? Answer: Q. LIh9: Select at low magnification the whole section and copy-paste it to the upper frame of this PPT slide. Q. LIh10: Select and magnify a B-cell follicle without a distinct germinal centre. Copy-paste it to your PPT. Q. LIh11: Select and magnify a secundary B-cell follicle (with a germinal centre). Copy-paste it to your PPT. Q. LIh12: Which cells in the germinal centre are very dark brown stained? Q. LIh13: What is the name of the follicle area around the germinal centre? Q. LIh14: What is the staining pattern of an individual B cell at higher magnification (10-20 x) and why is that? -> After answering the questions go back to the website and continue with part 6D: Lymph nodes; Immunohistology ‘T-cell (TCR) staining’

‘T-cell (TCR) staining’ Part 6D: Lymph nodes; Immunohistology ‘T-cell (TCR) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh15: Select at low magnification the whole section and copy-paste it to the upper frame of your PPT. Copy-paste the low magnification of the B-cell staining of the previous PPT slide to the lower frame of this one. Now compare the 2 sections. stained for T-cells with that stained for B-cells. You can easily recognize that the T-cell and B-cell areas are complementary to each other. Q. LIh16: What is the term for the T-cell area in the lymph node? Answer: Q. LIh17: Are T- and B-cells only present in their own area ? -> After answering the questions go back to the website and continue with part 6E: Lymph nodes; Immunohistology ‘T-helper cell (CD4) staining’

Part 6E: Lymph nodes; Immunohistology ‘T-helper cell (CD4) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh18: What is the way by which T-helper cells give help to other cells in the immune response? Answer: Q. LIh19: Select a T cell area and magnify 10x. Copy-paste the area to your PPT -> After answering the questions go back to the website and continue with part 6F: Lymph nodes; Immunohistology ‘T-cytotoxic cell (CD8) staining’ Part 6F: Lymph nodes; Immunohistology ‘T-cytotoxic cell (CD8) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: .Q. LIh21: Select the same T cell area as you did for the T-helper cells and magnify 10x.Copy-paste this area to your PPT. Q. LIh22: When you compare the shot of T-cytotoxic cells with that of T-helper cells, which of the two cell types is the highest in number in the T-cell area? What is the ratio of T-helper/T-cytotoxic cells in the T-cell area? Answer: -> After answering the questions go back to the website and continue with part 6G: Lymph nodes; Immunohistology ‘HEV (endothelium) staining’

‘HEV (endothelium) staining’ Part 6G: Lymph nodes; Immunohistology ‘HEV (endothelium) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh23: In which area of the lymph node are HEV present? Answer:  Q. LIh24: What is the function of an HEV? Answer: Q. LIh 25: Which specific molecules are present on the surface of the high-endothelial cells to allow this function? Answer:  Q. LIh26: To which cell type do the blue nuclei belong that you can see in lumen of the HEV? Answer: Q. LIh 27: Select and magnify 10-20x an HEV with the characteristic cuboidal endothelial cells and the ‘string of beads’ appearance. Copy-paste it to your PPT. Q. LIh 28: Select and magnify also10-20x a non-HEV vessel with characteristic flat endothelium. Copy-paste it to your PPT. -> After answering the questions go back to the website and continue with part 6H: Lymph nodes; Immunohistology ‘Dendritic cell (MHC-class II) staining’

‘Dendritic cell (MHC-class II) staining’ (1) Part 6H: Lymph nodes; Immunohistology ‘Dendritic cell (MHC-class II) staining’ (1) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh 29: In which areas of the lymph node do you find MHC-class II positive cells ? Answer:  Q. LIh30: Which cells in these areas do express MHC-class II molecules? Answer: Q. LIh31: Which T-cell type gets antigen presented by MHC-class II molecules? Q. LIh32: Which of these cell types is the most ‘professional’ APC? In which area is this cell present? Q. LIh33: In which area is this cell present? Q. LIh34: What is the function of the specific morphology of these cells?

‘Dendritic cell (MHC-class II) staining’ (2) Part 6H: Lymph nodes; Immunohistology ‘Dendritic cell (MHC-class II) staining’ (2) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh28: Copy-paste a low magnification of the section to your PPT. Q. LIh35: Select and magnify a T-cell area 10-20x and search for 1 or more dendritic cells (DCs) with the characteristic morphology (irregular shape with extensions between surrounding T-cells). Copy paste the selection to your PPT -> After answering the questions go back to the website and continue with part 6I: Lymph nodes; Immunohistology ‘Plasma cell (intracellular Ig) staining’

‘Plasma cell (intracellular Ig) staining’ Part 6I: Lymph nodes; Immunohistology ‘Plasma cell (intracellular Ig) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. LIh 36: In which area of the lymph node are plasma blast/cells formed ? Answer: Q. LIh 37: In which area of the lymph node do you find most mature plasma cells? Explain why this is not the area in which they are formed? Q. LIh 38: Select, and copy-paste a low magnification of a section part with cortex and medulla. Q. LIh 39: Select, magnify high (20x), and copy-paste a medulla area showing some plasma cells. Q. LIh 40: What is the light spot that can be distinguished in the brown plasma cells? Answer: -> After answering the questions of part 6I please continue with part 7: Spleen General histology

Part 7: Spleen; General histology (1) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. S1: Make a screenshot of the whole section and copy-paste it in the frame .

Part 7: Spleen; General histology (2) -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. S2: Describe the routing of blood that flows through to the spleen starting from the artery that enters the spleen and ending with the vein that leaves the spleen. Answer: Q. S3: Do you find HEVs in the T-cell areas of the splenic white pulp? Answer: Q. S4: Select and magnify a spleen area with red pulp and white pulp. Copy/paste it to your PPT, and indicate both areas. Q. S5: Select and magnify a white pulp area in which the central artery/arteriole, the PALS, a follicle and marginal zone can be identified. Copy/paste it to your PPT, and indicate each element. -> After answering the questions go back to the website and continue with part 8: Spleen; Immunohistology

Part 8A: Spleen; Immunohistology ‘Reticulum staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh1: Select and magnify a red pulp area, copy-paste it to your PPT Q. SIh2: Select and magnify a white pulp area, copy-paste it to your PPT. Q. SIh3: Select, magnify and copy-paste an inner PALS area showing the stained connective tissue of the central artery and surrounding connective tissue strands. -> After answering the questions go back to the website and continue with part 8B: Spleen; Immunohistology ‘Macrophage staining’

‘Macrophage staining’ Part 8B: Spleen; Immunohistology ‘Macrophage staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh4: In which area of the spleen do you find most scavenger macrophages ? Answer: Q. SIh5: Select and magnify an area with red and white pulp, copy-paste it to your PPT. Indicate what is the white pulp. Q. SIh6: Does the marginal zone contain scavenger macrophages ? -> After answering the questions go back to the website and continue with part 8C: Spleen; Immunohistology ‘B-cell (IgM) staining’

‘B-cell (IgM) staining’ Part 8C: Spleen; Immunohistology ‘B-cell (IgM) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh7: Do you find B-cells in the red pulp ? Answer: Q. SIh8: Select and magnify an area with red and white pulp, copy-paste it to your PPT. Indicate what is the white pulp. Q. SIh9: Search in the white pulp areas for a B-cell follicle with a germinal centre. Magnify and paste it to your PPT. -> After answering the questions go back to the website and continue with part 8D: Spleen; Immunohistology ‘Endothelium staining’

‘Endothelium staining’ Part 8D: Spleen; Immunohistology ‘Endothelium staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh11: Select and magnify (5x) a red pulp area in which you can recognize the stained sinuses. Q. SIh12: Select and magnify (5-10x) a white pulp area in which you can recognize the central arteriole of the PALS. -> After answering the questions go back to the website and continue with part 8E: Lymph nodes; Immunohistology ‘T-cell (TCR) staining’

‘T-cell (TCR) staining’ Part 8E: Spleen; Immunohistology ‘T-cell (TCR) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh13: Do you find T-cells in the red pulp ? Answer: Q. SIh14: Select and magnify (2,5x) a white pulp area, copy-paste it to your PPT. Indicate what is the white pulp. Q. SIh15: How is in the white pulp the area called that contains the T-cells ? -> After answering the questions go back to the website and continue with part 8F: Lymph nodes; Immunohistology ‘T-helper cell (CD4) staining’

Part 8F: Spleen; Immunohistology ‘T-helper cell (CD4) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh16: Select and magnify (5x) a white pulp area, copy-paste it to your PPT. -> After answering the questions go back to the website and continue with part 8G: Spleen; Immunohistology ‘T-cytotoxic cell (CD8) staining’ Part 8G: Spleen; Immunohistology ‘T-cytotoxic cell (CD8) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh17: Select and magnify (5x) the same white pulp area as in the section before, copy-paste it to your PPT.   Q. SIh18: Compare the CD4 and CD8 staining in your PPT. Is CD4 or CD8 the largest population of T-cells in the PALS ? Is this different as for lymph nodes? Answer: -> After answering the questions go back to the website and continue with part 8H: Spleen; Immunohistology ‘Dendritic cell (MHC-class II) staining’

‘Dendritic cell (MHC-class II) staining’ Part 8H: Spleen; Immunohistology ‘Dendritic cell (MHC-class II) staining’ -> Answer the questions below, use the virtual tissue section and the advised background information. Tasks and questions: Q. SIh19: In which areas of the white pulp do you find stained antigen presenting cells (APC) ? Answer: Q. SIh20: What is the specific cell type present in the T-cell area (inner PALS)? To which T-cell type is this APC presenting antigen by the MHC-class II molecule ? Q. SIh21: Select and magnify 10-20x an inner PALS area in which the characteristic morphology of the APC is shown. Copy-paste it to your PPT. -> This was the last part of the program. You may close your PPT and end the program or go back to a previous part of the program.