diff --git a/assays/S1_A2_microscopy_hyphae/protocols/Calcofluor white staining .md b/assays/S1_A2_microscopy_hyphae/protocols/Calcofluor white staining .md index 822d477f00b23e589d3a43bb30f9bfbdeac8f5ca..307e5bde5889d248e10c2e307d0365886dc21534 100644 --- a/assays/S1_A2_microscopy_hyphae/protocols/Calcofluor white staining .md +++ b/assays/S1_A2_microscopy_hyphae/protocols/Calcofluor white staining .md @@ -1 +1,2 @@ +# Calcofluor white staining To visualize basal septa and empty sections, 1 ml of cell culture was stained with 1 μl of Calcofluor white staining solution (2 mg/ml; CFW) directly before microscopy with a DAPI filter set \ No newline at end of file diff --git a/assays/S1_A2_microscopy_hyphae/protocols/Growth_condition.md b/assays/S1_A2_microscopy_hyphae/protocols/Growth_condition.md index 0862226d01f590fcdb1feffb193c89daa5bf0720..02885e62e851d369b5b04a3b6223ded8d81777f0 100644 --- a/assays/S1_A2_microscopy_hyphae/protocols/Growth_condition.md +++ b/assays/S1_A2_microscopy_hyphae/protocols/Growth_condition.md @@ -1 +1,2 @@ +# Growth conditions For microscopic analysis, 20 ml of yeast cells were grown in CM medium (1% glucose) to an OD600 of 0.5 Hyphal cells were induced by shifting 20 ml cell culture from CM medium to NM medium (supplemented either with 1% glucose or 1% arabinose) for 6, 9, 10, and 12 hours. \ No newline at end of file diff --git a/assays/S1_A2_microscopy_hyphae/protocols/measurements.md b/assays/S1_A2_microscopy_hyphae/protocols/measurements.md new file mode 100644 index 0000000000000000000000000000000000000000..29b17bed7a52efef4b8493e703f2bdcefeaf9037 --- /dev/null +++ b/assays/S1_A2_microscopy_hyphae/protocols/measurements.md @@ -0,0 +1,2 @@ +# measurements +The hyphal length was determined by measuring the distance between the tip and the basal pole without including the empty sections. Hyphal width was calculated by taking the average thickness measured at three different locations within each hypha.Hyphal cells with empty sections were visualized using Calcofluor White (CFW) and were scored manually. For each experiment, more than 100 cells were counted per strain. \ No newline at end of file diff --git a/assays/S1_A2_microscopy_hyphae/protocols/microscopy.md b/assays/S1_A2_microscopy_hyphae/protocols/microscopy.md index 22985f28899b3ee6836f035fbd1dacad05251098..9156ed69d82c86f62da0fad1833a0a2a1370b529 100644 --- a/assays/S1_A2_microscopy_hyphae/protocols/microscopy.md +++ b/assays/S1_A2_microscopy_hyphae/protocols/microscopy.md @@ -1,2 +1,3 @@ +# microscopy All images were acquired using laser-based epifluorescence-microscopy, Zeiss Axio Observer.Z1 (Oberkochen, Germany) and processed using the MetaMorph software package (version 7.7.0.0, Molecular Devices, Seattle, IL, USA). The hyphal length was determined by measuring the distance between the tip and the basal pole without including the empty sections. Hyphal width was calculated by taking the average thickness measured at three different locations within each hypha.Hyphal cells with empty sections were visualized using Calcofluor White (CFW) and were scored manually. For each experiment, more than 100 cells were counted per strain. \ No newline at end of file