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zivy committed Nov 28, 2023
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| Making 1 mg/mL of LiBH<sub>4</sub> [[video](https://www.youtube.com/watch?v=-MH_aLEMyF8)]. | {::nomarkdown}<details ><summary>Dissolve LiBH<sub>4</sub> (STREM...</summary><p>Dissolve LiBH<sub>4</sub> (STREM Chemicals, cat no. 93-0397; purchase in 1 g aliquots) into diH<sub>2</sub>O (Quality Biological, cat no. 351-029-101) and allow to sit at RT for 10 minutes until the formation of bubbles occurs (not shown in video). Pass the solution through a 0.22 &mu;m syringe filter (Millipore Sigma, cat no. SLGSM33SS) attached to a 20 mL syringe (EXEL Int., cat no. 26280) to remove any impurities prior to use (not shown).<br><br>Always prepare LiBH<sub>4</sub> solution in a chemical fume hood with appropriate personal protective equipment (PPE). Mixing LiBH<sub>4</sub> with diH<sub>2</sub>O will generate hydrogen gas which is highly flammable. For this reason, we always work with small amounts of LiBH<sub>4</sub> relative to the amount of water initially added. We do not mix >10 mg of LiBH<sub>4</sub> with diH<sub>2</sub>O. This amount can be quickly diluted to 1 mg/mL with volumes handled by standard serological pipets/pipet-aids. Always wrap LiBH<sub>4</sub> stock with parafilm (suggest Bemis Company Inc., cat no. S37440) and store in the presence of desiccant. We recommend replacing LiBH<sub>4</sub> every 4 weeks as repeated exposure to moisture/air has been observed to reduce fluorophore inactivation efficacy. Be sure to follow institutional guidelines for proper disposal of hazardous chemicals.<br><br>The formation of bubbles indicates the solution is ready for filtration and subsequent use. For best results, use solution immediately after filtration. We have noted effective fluorophore inactivation up to 4 hours after initial dissolution of LiBH<sub>4</sub>.</p></details>{:/} |
| Coating slide with chrome alum-gelatin adhesive [[video](https://www.youtube.com/watch?v=ksNR3gsl5rg)]. | {::nomarkdown}<details ><summary>Coat a glass...</summary><p>Coat a glass slide by adding 15 &mu;L chrome alum-gelatin to one side. Spread evenly using the edge of a cover slip or a separate glass slide via the blood smear technique. Note, it may require multiple spreading passages with the slide to ensure even coating of chrome alum-gelatin without streaks. If chrome alum-gelatin pools on the slide, aspirate excess fluid and respread as these accumulations can result in autofluorescence artifacts during image acquisition. Be mindful of stated expiration date on chrome alum-gelatin.<br><br>For best results, prepare chrome alum-gelatin coated slides freshly (i.e., the day of tissue sectioning) or no more than 7 days prior to tissue sectioning. If chrome alum-gelatin coated slides are prepared ahead of sectioning, store at room temperature (do not freeze as this will compromise the adhesive properties).<br><br>Dry coated slide in oven at 60°C for 1 hour.<br><br>We purchase chrome alum-gelatin from Newcomer Supply, but recipes are available online if this item does not ship to your country (e.g. [this recipe](https://www.laboratorynotes.com/preparation-of-chrome-alum-containing-gelatin-solution-for-preparation-of-coated-slides-for-histological-tissue-sections/))</p></details>{:/} |
| Mounting tissue and applying coverslip [[video](https://www.youtube.com/watch?v=tqHlrSmsG_8)]. | {::nomarkdown}<details ><summary>Remove as much...</summary><p>Remove as much PBS as possible without drying out tissues. Quickly add the minimum amount of Fluoromount-G mounting medium necessary to completely cover each tissue section. Gently cover with a coverslip. We typically use 10 – 40 &mu;L per tissue section. Ensure there are no bubbles on or near tissue.</p></details>{:/} |
| XTRegisterSameChannel - SimpleITK Imaris Python Extension [[video](https://www.youtube.com/watch?v=rrCajI8jroE)]. | {::nomarkdown}<details ><summary>This video illustrates...</summary><p>This video illustrates how to use the XTRegisterSameChannel SimpleITK Imaris (Oxford Instruments) Python Extension - registration of 2D or 3D images that share a common channel (correlation based affine alignment).</p></details>{:/} |
| XTRegisterSameChannel - SimpleITK Imaris Python Extension [[video](https://www.youtube.com/watch?v=rrCajI8jroE)]. | {::nomarkdown}<details ><summary>This video illustrates...</summary><p>This video illustrates how to use the XTRegisterSameChannel SimpleITK Imaris (Oxford Instruments) Python Extension - registration of 2D or 3D images that share a common channel (correlation based affine alignment).</p></details>{:/} |
| Leica LAS X Navigator: Focus Map [[video](https://www.youtube.com/watch?v=SbDDzo8dPZo)]. | {::nomarkdown}<details ><summary>This is a...</summary><p>This is a video tutorial on how to set up a focus map in LAS X Navigator (v. 3.6.0 Widefield), which is recommended for tissue sections or samples that are continuous.</p></details>{:/} |
| Leica LAS X: Hardware Autofocus (AFC) [[video](https://www.youtube.com/watch?v=3uq42XefHhY)]. | {::nomarkdown}<details ><summary>How to use...</summary><p>How to use Adaptive Focus Control (AFC) in your Navigator experiments, if your DMi8 microscope is equipped with AFC.</p></details>{:/} |
| Leica LAS X Linked Shading: Fluorescence [[video](https://www.youtube.com/watch?v=9l_DmMmAbHk)]. | {::nomarkdown}<details ><summary>A video tutorial...</summary><p>A video tutorial on how to do Linked Shading (shading correction) in LAS X 3.6 (widefield) with fluorescent images on a monochrome camera.</p></details>{:/} |
| Leica LAS X THUNDER Tutorial [[video](https://www.youtube.com/watch?v=36HqrFyy0fQ)]. | {::nomarkdown}<details ><summary>This video shows...</summary><p>This video shows how to "THUNDER" images in LAS X on-the-fly or post-acquisition with the default settings. (v. 3.7.1 - Widefield). This is only available on THUNDER Imagers.</p></details>{:/} |
| Leica LAS X Software Experiment Setup: Z-stack [[video](https://www.youtube.com/watch?v=dTMF01cO7lI)]. | {::nomarkdown}<details ><summary>How to set...</summary><p>How to set up a Z-stack in LAS X v 3.3 (widefield). |
| | Note: the example in this video was for a data set that was intended for deconvolution, so it goes slightly beyond the focus on either end. The usual recommendation is to stay mostly in focus for all of your Z planes, so that subsequent 2D projections (such as a maximum intensity projection or extended depth of focus projection) will be clearer.</p></details>{:/} |

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