Session 8

Advanced and developmental microscopy

Chat Transcript

00:20:45 Liisa Hirvonen: Thanks Gavin, really beautiful images, looks like a very versatile instrument! Will you get a demo system to Australia?
00:24:13 Gavin Symonds: Hello Liisa. Yes, the Elyra 7 is a very versatlie instrument. We have Elyra 7 instruments installed in Australia, although due to COVID this makes things more complicated. But we can discuss getting you some exposure to the Elyra 7 instrument.
00:28:35 Gavin Symonds: Also Liisa, ZEISS will be hosting an Elyra 7 workshop next Wednesday afternoon, that everyone is welcome to attend. Please visit the ZEISS virtual booth on the LMA website for further details.
00:29:13 Liisa Hirvonen: Thanks Gavin! 🙂
00:31:10 Louise Cole: Please remember to post your questions in the chat
00:42:28 Renee Whan: @chao, does the lattice beam alignment occur predominantly automatically or manually?
00:42:54 Senthil Arumugam: @Chao – are multiple wavelength sequential through the SLM?
00:43:04 Liisa Hirvonen: Bruker/Luxendo: very nice developments! But looks like you have lots of different microscopes for different applications. What would you recommend for a core facility that needs to cater for lots of different researchers, samples and applications?
00:46:57 Louise Cole: @gavin Does SIM2 allows live cell imaging on the fly. It was not clear to me.
00:47:47 Louise Cole: @carola I am interested in how deep one can image with the Easy 3D STED mode
00:47:50 Andleeb: Carola can you please share registration link to join STED workshop?
00:48:57 Chao Zheng: @Renee, the lattice beam alignment on InVi lattice is a one time hardware alignment during installation to correct any optical shifts/movement during shipments, once it is aligned, during the daily usage, all alignments are motorized, one can accurately align the system in software within a few minutes.
00:53:11 Louise Cole: @Zheng Chao you have a wide-range of LISH microscopes available – how important is the size of the microscope footprint when developing commercial LISH technology? Do you think we will ever to get the point where these microscopes are portable and could be used in the field or clinical settings
00:55:08 Carola Thoni: We were running the workshop already at the 3rd of August. But we recorded all. Here is the link to the recording:
00:55:30 Chao Zheng: @Senthil, InVi lattice supports simultaneous dual channel detection, however, for lattice imaging, we advise user to image multichannel sequentially, because same lattice pattern does not give best image quality for different wavelength. Slight tweak of lattice beam parameter is needed when switching to different wavelength. However, instead of switching mechanical filter wheels, we will support fast switching of excitation lasers to minimize acquisition time difference between channels. simultaneous multi channel acquisition using Gaussian/Bessel beam has no issue.
00:56:05 Carola Thoni: Here is the link to the workshop recording:
00:56:24 Andleeb: Thank You @Carola
00:56:28 Gavin Symonds: @Louise. Direct or External processing is available that can be peformed on the Lattice SIM or Apotome SIM data as it is being acquired using an offline storage computer. SIM^2 processing can be peformed on Lattice SIM image data, as well as Apotome SIM data. And the SIM^2 can be peformed when combined with other processing methods such as Burst mode and Leap mode.
00:56:28 Senthil Arumugam: @Chao, Thanks
00:57:13 Louise Cole: Thanks @gavin
00:58:06 Chao Zheng: @Liise Yes, indeed. it is hard to suggest one system for all possible applications, because it can range from single layer cell to whole cleared organs/animals. However, MuVi-SPIM do cover the widest range of applications, including organoids, fishes, c-elegans, plants, and cleared samples up to 15mm.
00:58:35 Liisa Hirvonen: Thanks Chao! 🙂
00:58:49 Louise Cole: That is good to know – thanks @zheng chao
00:59:06 Renee Whan: @Ying, im wondering what stain you used to look at your F-actin in your experiments. What you think the role of actin is specifically related to necroptosis?
01:03:02 Carola Thoni: Combining Easy3D STED, Adaptive Illumination and Deformable Mirror we can image more than 100 µm in tissue. I have seen 120 µm deep STED imaging in muscle tissue with the Abberior Instruments Facility Line STED system we just installed in Auckland.
01:04:24 Louise Cole: That’s great – thanks @Caarola
01:05:46 Renee Whan: @ash, segmenting each microtubule is not particularly easy when do tracing, could you tell us a little about how SIFNE works?
01:09:30 Chao Zheng: @Louise the size of footprint is not that critical for research labs, normally space is not the biggest constrains in labs, instead, user friendly imaging and maintenance are. It is an very interesting idea to make lightsheet microscope portable, and it does make sense because it is one of the best microscope techniques to acquire 3D long time lapse images. and the latest development is to combine lightsheet with light field to have video-frame-rate 3D image acquisitions. the smallest lightsheet microscope is roughly of luggage size now, it will take more efforts to make it really portable in the future.
01:11:38 Renee Whan: MORE COFFEE
01:11:55 Samantha Stehbens: @ashley- I may have missed it. Do you have insight to the mechanism of curvature in the polymer in response to the drug? Are you planning to test other microtubule drugs?
01:16:09 Oleks Chernyavskiy: @Louise: There is The Flamingo Project describing portable LS microscope by Jan Huisken: “Designed as a portable, shareable light sheet microscope, Flamingo essentially shrinks a tabletop-sized technology down to the weight and dimensions of a suitcase.”
01:19:52 Louise Cole: Thank you @Oleks
01:27:35 Harrison York: @ying you mentioned that MLKL is actively trafficked to the PM and that it was dependent on the cytoskeleton. Have you investigated which motor proteins might be involved or through which compartments it is trafficked?
01:32:17 radek: @ying have you looked at involvement of nuclear actin in seen phenotype, or is it purely a cytoplasmic portion of actin that is required form MLKL foci formation?
01:34:11 Samantha Stehbens: Thank-you Ashley!