# Probes & Prep **URL:** https://forum.microlist.org/c/probes-prep/9.md?page=2 [Latest](https://forum.microlist.org/latest.md) · [Categories](https://forum.microlist.org/categories.md) · [Tags](https://forum.microlist.org/tags.md) **Page:** 3 --- ## [Re-usable live cell chamber](https://forum.microlist.org/t/re-usable-live-cell-chamber/1143)
**Author:** [@WillGiang](https://forum.microlist.org/u/WillGiang)\ **Replies:** 4\ **Last updated:** [April 1, 2021, 9:24am UTC](https://forum.microlist.org/t/re-usable-live-cell-chamber/1143 "2021-04-01T09:24:41Z")
Hi, Do people have a favorite re-usable live cell chamber? The Attofluor checks all of my boxes, but I’m wondering what else is out there. Thanks, Will --- ## [Experience with 440nm excitable fluorophores? Brilliant violet 480, Biotium Cf430, not working](https://forum.microlist.org/t/experience-with-440nm-excitable-fluorophores-brilliant-violet-480-biotium-cf430-not-working/1331)
**Author:** [@mssher07](https://forum.microlist.org/u/mssher07)\ **Replies:** 2\ **Last updated:** [March 16, 2021, 3:00pm UTC](https://forum.microlist.org/t/experience-with-440nm-excitable-fluorophores-brilliant-violet-480-biotium-cf430-not-working/1331 "2021-03-16T15:00:19Z")
I have a very strange problem. We have a confocal microscope with a laser launch partly optimized for live cell imaging with CFP, so we have a 440nm laser instead of the usual 405nm. We know this laser works because my l… --- ## [Recommendations for fluorescent nano-beads for PSF confocal](https://forum.microlist.org/t/recommendations-for-fluorescent-nano-beads-for-psf-confocal/1275)
**Author:** [@NachoDave](https://forum.microlist.org/u/NachoDave)\ **Replies:** 7\ **Last updated:** [February 18, 2021, 11:40pm UTC](https://forum.microlist.org/t/recommendations-for-fluorescent-nano-beads-for-psf-confocal/1275 "2021-02-18T23:40:19Z")
Hi, We have an INCELL 6000 analyzer, which has confocal capabilities, using a line scanning confocal, which appears to be somewhere between wide field and confocal. Images are quite blurry, especially in the z-plane. W… --- ## [Unspecific signals in fluorescence staining of neutrophil extracellular traps](https://forum.microlist.org/t/unspecific-signals-in-fluorescence-staining-of-neutrophil-extracellular-traps/1129)
**Author:** [@Francesca](https://forum.microlist.org/u/Francesca)\ **Replies:** 2\ **Last updated:** [February 12, 2021, 6:12pm UTC](https://forum.microlist.org/t/unspecific-signals-in-fluorescence-staining-of-neutrophil-extracellular-traps/1129 "2021-02-12T18:12:29Z")
Dear all, I am trying to stain RNA in neutrophil extracellular traps by using appropriate primary antibodies and corresponding secondary antibodies (polyclonal from chicken with Alexa Flour 594 or 647 tag). I am facing … --- ## [Beads for channel alignment](https://forum.microlist.org/t/beads-for-channel-alignment/1197)
**Author:** [@sri1989vathsan](https://forum.microlist.org/u/sri1989vathsan)\ **Replies:** 2\ **Last updated:** [February 11, 2021, 7:53pm UTC](https://forum.microlist.org/t/beads-for-channel-alignment/1197 "2021-02-11T19:53:23Z")
I am looking for a set of 100nm beads that I can use for aligning different channels. I want to add these beads to my sample (as people do with EM) and image across the green (488 nm excitation), orange (561nm excitatio… --- ## [Coverslip holder](https://forum.microlist.org/t/coverslip-holder/1272)
**Author:** [@EricGriffis](https://forum.microlist.org/u/EricGriffis)\ **Replies:** 1\ **Last updated:** [February 10, 2021, 12:34pm UTC](https://forum.microlist.org/t/coverslip-holder/1272 "2021-02-10T12:34:38Z")
Hi All, Does anyone know if anyone makes a drop in adapter that would allow me to place a 24x40mm or 24x50mm coverslip into a form that is 25x75mm that would then fit well into an stage top incubator? I have someone who… --- ## [Live Golgi marker](https://forum.microlist.org/t/live-golgi-marker/1281)
**Author:** [@mapalmer](https://forum.microlist.org/u/mapalmer)\ **Replies:** 2\ **Last updated:** [February 8, 2021, 12:59pm UTC](https://forum.microlist.org/t/live-golgi-marker/1281 "2021-02-08T12:59:57Z")
Hi, I was wondering if anyone has any recommendations for a live Golgi marker please? I will be working with primary human macrophages (non-proliferative) in addition to using BODIPY Cholesterol (TopFluor) 495 nm / 507… --- ## [Getting HEK293 cells to adhere to glass chamber slides for ICC](https://forum.microlist.org/t/getting-hek293-cells-to-adhere-to-glass-chamber-slides-for-icc/1263)
**Author:** [@achamess](https://forum.microlist.org/u/achamess)\ **Replies:** 2\ **Last updated:** [January 27, 2021, 3:34pm UTC](https://forum.microlist.org/t/getting-hek293-cells-to-adhere-to-glass-chamber-slides-for-icc/1263 "2021-01-27T15:34:53Z")
I want to perform immunocytochemistry on transfected HEK cells. I’ve been getting them to grow and adhere to Ibidi 12 well glass slides while they’re growing. I can do media changes and they stay. But once I take the out… --- ## [Combine brightfield and fluorescence](https://forum.microlist.org/t/combine-brightfield-and-fluorescence/1216)
**Author:** [@mariok](https://forum.microlist.org/u/mariok)\ **Replies:** 4\ **Last updated:** [December 3, 2020, 8:28am UTC](https://forum.microlist.org/t/combine-brightfield-and-fluorescence/1216 "2020-12-03T08:28:15Z")
Hello, our slide scanner has troubles detecting small tissue biopsies using the darkfield preview. Hence, I would like to combine a brightfield counterstain (tissue detection) with FITC IF imaging (specific antibody) … --- ## [STED Nanoruler and/or Calibration](https://forum.microlist.org/t/sted-nanoruler-and-or-calibration/1189)
**Author:** [@nranthony](https://forum.microlist.org/u/nranthony)\ **Replies:** 3\ **Last updated:** [November 18, 2020, 6:04pm UTC](https://forum.microlist.org/t/sted-nanoruler-and-or-calibration/1189 "2020-11-18T18:04:02Z")
Hi all, I was wondering if anybody knows of a good option for calibrating STED resolution? I’m aware of (and their 3D option, but not sure how that’s setup… are they all vertical maybe?) Are there others? I als… --- ## [Database for Photoswitchable FPs and dyes?](https://forum.microlist.org/t/database-for-photoswitchable-fps-and-dyes/1182)
**Author:** [@NC69390445](https://forum.microlist.org/u/NC69390445)\ **Replies:** 1\ **Last updated:** [October 28, 2020, 1:04pm UTC](https://forum.microlist.org/t/database-for-photoswitchable-fps-and-dyes/1182 "2020-10-28T13:04:21Z")
Is there a database for dyes and FPs used in SMLM? --- ## [Slowfade glass softset mounting media stoarge](https://forum.microlist.org/t/slowfade-glass-softset-mounting-media-stoarge/1172)
**Author:** [@manerotoni](https://forum.microlist.org/u/manerotoni)\ **Replies:** 0\ **Last updated:** [October 14, 2020, 2:19pm UTC](https://forum.microlist.org/t/slowfade-glass-softset-mounting-media-stoarge/1172 "2020-10-14T14:19:29Z")
Dear microscopist, I was wondering if you have any suggestions on how much one can thaw-cycle the slowfade glass mounting media Should one conserve it with silica in the fridge or always thaw it when needed? Somehow t… --- ## [Ibidi polymer coverslip](https://forum.microlist.org/t/ibidi-polymer-coverslip/1118)
**Author:** [@bioimage-analysis](https://forum.microlist.org/u/bioimage-analysis)\ **Replies:** 4\ **Last updated:** [September 2, 2020, 10:03pm UTC](https://forum.microlist.org/t/ibidi-polymer-coverslip/1118 "2020-09-02T22:03:16Z")
Hi everyone, The Nikon imaging center at Harvard website (@jennifer, @talley) mentioned issue with the ibidi polymer coverslip in combination with the Nikon oil. I wonder if it’s still an issue and if anyone had similar… --- ## [Snap-Cell 430 Advice](https://forum.microlist.org/t/snap-cell-430-advice/1109)
**Author:** [@EricGriffis](https://forum.microlist.org/u/EricGriffis)\ **Replies:** 0\ **Last updated:** [August 17, 2020, 8:47pm UTC](https://forum.microlist.org/t/snap-cell-430-advice/1109 "2020-08-17T20:47:59Z")
Does anyone have experience using this probe in either human cells or S. cerevisiae? We have a standard CSU-X1 with a 405nm line from a MLC400B and an emission filter wheel. I’m debating between recommending this or LSS-… --- ## [Expansion microscopy on lamina](https://forum.microlist.org/t/expansion-microscopy-on-lamina/1098)
**Author:** [@LPUoO](https://forum.microlist.org/u/LPUoO)\ **Replies:** 1\ **Last updated:** [August 7, 2020, 11:38am UTC](https://forum.microlist.org/t/expansion-microscopy-on-lamina/1098 "2020-08-07T11:38:43Z")
Hi, Does anyone have experience with expansion microscopy on lamina ? I tried the protocol from Gambarotto, D.et al. (2018). Imaging cellular ultrastructures using expansion microscopy (U-ExM). Nature Methods . https:/… --- ## [mEos versions and other recommendations for point localization in tissue](https://forum.microlist.org/t/meos-versions-and-other-recommendations-for-point-localization-in-tissue/1087)
**Author:** [@lisacam7](https://forum.microlist.org/u/lisacam7)\ **Replies:** 0\ **Last updated:** [July 24, 2020, 1:50pm UTC](https://forum.microlist.org/t/meos-versions-and-other-recommendations-for-point-localization-in-tissue/1087 "2020-07-24T13:50:04Z")
Hello - would welcome suggestions and experience with mEos versions for point localization. I’m assisting a user in my core who wants to initially test for inclusion in grant application. Post-doc has expressed mEos4b-ta… --- ## [Working with genetically encoded Redox sensors?](https://forum.microlist.org/t/working-with-genetically-encoded-redox-sensors/1062)
**Author:** [@elou888](https://forum.microlist.org/u/elou888)\ **Replies:** 0\ **Last updated:** [July 2, 2020, 9:57am UTC](https://forum.microlist.org/t/working-with-genetically-encoded-redox-sensors/1062 "2020-07-02T09:57:19Z")
Does anyone work with genetically encoded redox sensors like roGFP fusion proteins? I’d like to share experiences about probe preparations, imaging, data processing. And, especially, which hardware do you work with? --- ## [How to send coverslips across the world?](https://forum.microlist.org/t/how-to-send-coverslips-across-the-world/993)
**Author:** [@LPUoO](https://forum.microlist.org/u/LPUoO)\ **Replies:** 2\ **Last updated:** [July 1, 2020, 2:35pm UTC](https://forum.microlist.org/t/how-to-send-coverslips-across-the-world/993 "2020-07-01T14:35:24Z")
Hi, Does any one have tips for sending 13 mm round coverslips with adhered cells that need to be stained in another lab ? Mount them with non hardening media, seal them with nail varnish, send and unmount on arrival ? … --- ## [Do cells adhere less to sonnicated coverslips?](https://forum.microlist.org/t/do-cells-adhere-less-to-sonnicated-coverslips/962)
**Author:** [@LPUoO](https://forum.microlist.org/u/LPUoO)\ **Replies:** 3\ **Last updated:** [May 20, 2020, 10:19am UTC](https://forum.microlist.org/t/do-cells-adhere-less-to-sonnicated-coverslips/962 "2020-05-20T10:19:20Z")
Hi, I found that cells adhere less well to sonicated coverslips (following this protocol for example https://nic.med.harvard.edu/resources/coverslips/) Do people that routinely clean coverslips in the way of the protoc… --- ## [Avoid meniscus in 96 Well-Plate](https://forum.microlist.org/t/avoid-meniscus-in-96-well-plate/798)
**Author:** [@NicoKiaru](https://forum.microlist.org/u/NicoKiaru)\ **Replies:** 9\ **Last updated:** [March 23, 2020, 2:50am UTC](https://forum.microlist.org/t/avoid-meniscus-in-96-well-plate/798 "2020-03-23T02:50:33Z")
Hello everybody, The meniscus in the wells of the 96-well plate really affects the image quality in transmission imaging (it’s like adding a lens before the sample…). Do you know of anything that could be used to reduce… --- ## [Homemade Mounting Media](https://forum.microlist.org/t/homemade-mounting-media/908)
**Author:** [@EricGriffis](https://forum.microlist.org/u/EricGriffis)\ **Replies:** 1\ **Last updated:** [March 11, 2020, 12:33pm UTC](https://forum.microlist.org/t/homemade-mounting-media/908 "2020-03-11T12:33:13Z")
Does anyone have a mounting media that they love for multi-well plates? We’ve been recommending the standard 75% glycerol + propyl gallate, but if there is a better one, we’d be happy to try it. Bonus points if it has a … --- ## [Mineral oil to prevent evaporation for long time course on live cells](https://forum.microlist.org/t/mineral-oil-to-prevent-evaporation-for-long-time-course-on-live-cells/899)
**Author:** [@LPUoO](https://forum.microlist.org/u/LPUoO)\ **Replies:** 8\ **Last updated:** [March 10, 2020, 4:46pm UTC](https://forum.microlist.org/t/mineral-oil-to-prevent-evaporation-for-long-time-course-on-live-cells/899 "2020-03-10T16:46:21Z")
Hi, Some report using mineral oil to cover the media during long time courses with live cells. My question is whether the oil is gas permeable ? Are there any papers comparing gas permeability of different mineral oil… --- ## [96-well plate lids](https://forum.microlist.org/t/96-well-plate-lids/878)
**Author:** [@EricGriffis](https://forum.microlist.org/u/EricGriffis)\ **Replies:** 7\ **Last updated:** [March 10, 2020, 4:44pm UTC](https://forum.microlist.org/t/96-well-plate-lids/878 "2020-03-10T16:44:44Z")
Does anyone have a lid that they like for 96-well plates that is DIC compatible. I have a user that wants to use our robotic plate loading system to monitor organoid growth over a period of several days. It would be idea… --- ## [Protect bottom of optical plates](https://forum.microlist.org/t/protect-bottom-of-optical-plates/875)
**Author:** [@LPUoO](https://forum.microlist.org/u/LPUoO)\ **Replies:** 5\ **Last updated:** [February 22, 2020, 1:42pm UTC](https://forum.microlist.org/t/protect-bottom-of-optical-plates/875 "2020-02-22T13:42:46Z")
Hi, I used to use optical 96 well plates from Ibidi and they had a protective film in the bottom that I would remove just before imaging. I currently use a brand that doesn’t have that protective film. And because of t… --- ## [Slicing tissue blocks](https://forum.microlist.org/t/slicing-tissue-blocks/870)
**Author:** [@Remi\_A](https://forum.microlist.org/u/Remi_A)\ **Replies:** 2\ **Last updated:** [February 14, 2020, 2:25pm UTC](https://forum.microlist.org/t/slicing-tissue-blocks/870 "2020-02-14T14:25:53Z")
I will appreciate if anyone could tip me about how best to position and make thin slices (4-5um) from tissue blocks when such tissue blocks are irregular in shape, without wasting these samples. For context, these blocks… --- ## [Non genetically encoded calcium sensor](https://forum.microlist.org/t/non-genetically-encoded-calcium-sensor/869)
**Author:** [@Nat\_Prunet](https://forum.microlist.org/u/Nat_Prunet)\ **Replies:** 0\ **Last updated:** [February 12, 2020, 1:24am UTC](https://forum.microlist.org/t/non-genetically-encoded-calcium-sensor/869 "2020-02-12T01:24:15Z")
Any recommendation for a non-genetically encoded ratiometric calcium sensor for live cell imaging in mice? One of our core’s users wanted to use Fura 2 but don’t have microscopes equipped for both 340/380nm excitation an… --- ## [Live Cell Imaging Media Options](https://forum.microlist.org/t/live-cell-imaging-media-options/848)
**Author:** [@WillGiang](https://forum.microlist.org/u/WillGiang)\ **Replies:** 6\ **Last updated:** [February 7, 2020, 3:33am UTC](https://forum.microlist.org/t/live-cell-imaging-media-options/848 "2020-02-07T03:33:34Z")
Hi all, Has anyone compared the live cell imaging media sold by Sigma with Invitrogen’s Live Cell Imaging Solution or Gibco’s FluoroBrite DMEM? Are there alternatives to consider? Thanks in advance, Will --- ## [Diptera/mosquito archived slide of curious view](https://forum.microlist.org/t/diptera-mosquito-archived-slide-of-curious-view/847)
**Author:** [@mmcgary44](https://forum.microlist.org/u/mmcgary44)\ **Replies:** 0\ **Last updated:** [February 2, 2020, 4:51am UTC](https://forum.microlist.org/t/diptera-mosquito-archived-slide-of-curious-view/847 "2020-02-02T04:51:08Z")
It has so chanced I have managed now 3 years of collection to slide…\*library. Diptera. mosquito, Walla Walla, county, WA. CONUS. At year one exemplar, \*unstained, light field survey and curious as to near mouth part,… --- ## [Fiducials in Single Molecule Imaging](https://forum.microlist.org/t/fiducials-in-single-molecule-imaging/534)
**Author:** [@JASlotman](https://forum.microlist.org/u/JASlotman)\ **Replies:** 11\ **Last updated:** [January 29, 2020, 11:13am UTC](https://forum.microlist.org/t/fiducials-in-single-molecule-imaging/534 "2020-01-29T11:13:01Z")
I was wondering in what ways do you deposit fiducials and what do you use as fiducials. We have tried fluorescent beads (tetra speck) and colloidal gold particles. But I am not really satisfied by both of them, while the… --- ## [DNA labelling for lightsheet microscope](https://forum.microlist.org/t/dna-labelling-for-lightsheet-microscope/718)
**Author:** [@Alex.h](https://forum.microlist.org/u/Alex.h)\ **Replies:** 3\ **Last updated:** [January 28, 2020, 3:47am UTC](https://forum.microlist.org/t/dna-labelling-for-lightsheet-microscope/718 "2020-01-28T03:47:13Z")
Dear all, We use different protocol to clear our organ like X-clarity, CUBIC, RapiClear etc… But we have some problem with the DNA Staining. I measure that we stained with DAPI +/- 600 µm. The majority of our samples ar… [Previous page](https://forum.microlist.org/c/probes-prep/9.md?page=1) [Next page](https://forum.microlist.org/c/probes-prep/9.md?page=3)