In the week ten laboratory class the mitochondrial networks of wild-type and p...
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In the week ten laboratory class the mitochondrial networks of wild-type and patient-derived fibroblasts were visualised using fluorescent staining and microscopy. The expected results for the wild-type fibroblasts (well #1) and patient-derived fibroblasts (well #2) are shown in Figure 1 and are available in the week ten real-time section on Moodle. Complete Tasks A-B below to upload and interpret your results.
Figure 1. Expected fluorescence staining patterns for wild-type (#1) and patient-derived (#2) fibroblasts labelled with Streptavidin AF594 (red), Phalloidin AF488 (green), and DAPI (blue).
A) Results Upload: Upload an overlay fluorescence image (DAPI + GFP + Texas Red) showing your results for both the wild-type fibroblasts (well #1) and the patient-derived fibroblasts (well #2). Present both images in a single file (Word or PowerPoint) and ensure that each image is clearly labelled with the cell line (wild-type or patient) and corresponding well number (#1 or #2).
B) Fluorescent Signals: If the fluorescent staining procedure was successful, fluorescent signals should be visible in all three channels (DAPI, GFP, and Texas Red), as shown in Figure 1. Examine your experimental results. Did you successfully detect fluorescent signals in both the wild-type and patient-derived fibroblasts for all three fluorescent dyes (Streptavidin AF594, Phalloidin AF488, and DAPI)? (YES/NO) (4 marks)
If YES, proceed to the next question. (Q3)
If NO, identify which fluorescent dye(s) did not successfully produce a signal and suggest one or more possible sources of error or procedural issues that could explain this result.