Sp8 white light laser confocal microscope

Manufactured by Leica

The SP8 white light laser confocal microscope is a high-performance imaging system designed for advanced biological and materials science research. It features a tunable white light laser source that provides continuous coverage of the visible spectrum, enabling flexible and precise excitation of a wide range of fluorophores. The microscope is equipped with a high-speed scanner and sensitive detectors, allowing for rapid image acquisition and high-resolution imaging of live and fixed samples.

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Lab products found in correlation

Levamisole, by Merck Group (1 mentions) Bdm 2 3 butanedione monoxime, by Merck Group (1 mentions) Nis element, by Nikon (1 mentions) Ht501128, by Merck Group (1 mentions) Fluormount g, by Electron Microscopy Sciences (1 mentions) Citrisolv, by Decon Labs (1 mentions) Alexa fluor 488, by Jackson ImmunoResearch (1 mentions)

Close spelling variants of this product

TCS SP8 X white light laser inverted confocal microscope SP8 X white light laser confocal microscope SP8 white light laser scanning confocal microscope Leica TCS SP8 X White Light Laser confocal microscope SP8 white laser microscope SP8 laser confocal microscope White light laser SP8 confocal microscope Laser confocal microscope Confocal SP8

8 protocols using sp8 white light laser confocal microscope

Visualizing Protein Synthesis Dynamics

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Example 27

HEK293T cells were treated with 15 μM of OPP for 30 min. After that, they were fixed with 3.7% PFA/PBS, washed with PBS, permeabilized with 0.5% Triton X-100 in PBS and washed again. Then, the cells were labeled for 20 min with either compound 23, 24, 40, 41 and picolyl azide conjugated to Alexa Fluor 488 and Alexa Fluor 594 dyes (4.5 μM), 0.25 mM, 0.5 mM, 1 mM copper sulfate with or without 0.5, 1, or 2 mM THPTA respectively. Concentration of ascorbic acid in all cases was 6 mM. The cells were washed with 0.5 mM EDTA, 2 mM sodium azde in PBS, stained with Hoechst 33342. Negative controls were cell treated treated with DMSO. Images were captured on Leica SP8 White Light Laser Confocal microscope.

US11639342B2. 1,3-dipolar cycloadditions, and Staudinger ligations for conjugating biomolecules using click chemistry (2023-05-02). Bioconjugate Technologies, LLC [US]. Inventors: Andrei Polukhtin [US], Peter J. An [US], Alexandra Kulyashova [US], Jakub M. Labedz [US], Valentin A. Rassadin [US], Nikita V. Savelyev [US], Kostiantyn Ziabrev [US].

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Fluorescence Imaging of Paralyzed C. elegans

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Degradation groups were created as described above. All images were taken using 96-h-old animals. Adult animals were anesthetized by being placed in 10 µl of 7.5 mM Levamisole (Sigma-Aldrich) and 0.225 M BDM (2,3-butanedione monoxime) (Sigma-Aldrich) on glass microscope slide containing a 2% agar pad. After animals were paralyzed, a 1.5 coverslip was placed on top the agar pad. A Leica SP8 white light laser confocal microscope and 63× oil immersion lens was used for imaging. Step size was 0.3 µm. GFP was excited using a 488 nm wavelength laser with emitted light collected through a 490-778 nm bandpass filter. mNeonGreen was excited using a 506 nm wavelength laser with emitted light collected through a 512-742 nm bandpass filter. For each condition, four to five animals were scored for the presence of GFP or mNeonGreen. Final figures were generated using ImageJ (National Institutes of Health, Bethesda, MD, USA).

Kepler L.D., McDiarmid T.A, & Rankin C.H. (2022). Rapid assessment of the temporal function and phenotypic reversibility of neurodevelopmental disorder risk genes in Caenorhabditis elegans. Disease Models & Mechanisms, 15(5), dmm049359.

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Immunofluorescence Analysis of Mitochondrial Morphology

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Cells were fixed for 10 minutes with 4% paraformaldehyde in 1X PBS (phosphate buffer saline), followed by permeabilization with 1X PBS-0.25% (v/v) Triton X-100. After blocking for 45 min with 5% BSA in 1X PBS/0.25% Triton X-100/, cells were incubated with the primary antibodies mouse anti-Myc (1:2000) and/or rabbit anti-HSP60 (1:3000). The secondary antibodies used were anti-mouse-Alexa Fluor 488 and anti-rabbit Alexa Fluor 555 or 594 (1:500). Nuclei were stained with DAPI (1 μg/ml). Images were acquired on a LEICA SP8 White Light Laser confocal microscope and were analyzed using Image J. Mitochondrial network morphology was analyzed using the Mitochondria Analyzer plugin (13 ), using the following settings: 2D threshold, mean with block size at 1.450 microns and C-value at 5.

Papadaki V., Erpapazoglou Z., Kokkori M., Rogalska M.E., Potiri M., Birladeanu A., Tsakiri E.N., Ashktorab H., Smoot D.T., Papanikolopoulou K., Samiotaki M, & Kafasla P. (2023). IQGAP1 mediates the communication between the nucleus and the mitochondria via NDUFS4 alternative splicing. NAR Cancer, 5(3), zcad046.

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Confocal Imaging of Embryonic Development

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Images were acquired on a Leica SP8 White Light laser confocal microscope. Image processing was completed using Nikon NIS-Elements multi-platform acquisition software with a 40×/1.10 Water objective. Fiji (ImageJ, V 2.0.0-rc-69/1.52p) was utilized to color DAPI channel to cyan, GFP color remained green. Confocal images are max projections of Z stacks taken 0.5 μm apart for a total of the embryo ~7–12 μm.

Hickey G.J., Wike C.L., Nie X., Guo Y., Tan M., Murphy P.J, & Cairns B.R. (2022). Establishment of developmental gene silencing by ordered polycomb complex recruitment in early zebrafish embryos. eLife, 11, e67738.

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Arabidopsis Leaf Transient Transformation

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Intact leaves of 3-week-old soil-grown Arabidopsis plants were bombarded with a plasmid for expression of GFP using microprojectile bombardment as previously described (52 (link)). Three biological replicates in duplicate were performed. Bombarded plants were kept in growth chambers for 24 hours before individual transformed cells were identified using a Leica SP8 White Light Laser confocal microscope using the 488-nm line of the argon ion laser for excitation and emission collected between 505 and 530 nm. Layers of cells containing trafficked GFP surrounding the transformed cells were counted. Images were collected with a 25× 0.95 NA water immersion lens.

Lim G.H., Liu H., Yu K., Liu R., Shine M.B., Fernandez J., Burch-Smith T., Mobley J.K., McLetchie N., Kachroo A, & Kachroo P. (2020). The plant cuticle regulates apoplastic transport of salicylic acid during systemic acquired resistance. Science Advances, 6(19), eaaz0478.

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Formalin-Fixed Tissue Histology

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Freshly dissected tissues were fixed overnight at 4°C in 10% neutral buffered formalin (Sigma; HT501128). Fixed tissues were transferred to 70% ethanol for storage at 4 °C. Processing for histology followed standard protocols with paraffin embedding, sectioning at 5 μm thickness, baking for 1 h at 55 °C, and deparaffinizing with CitriSolv (Decon Labs, 89426–268) and rehydration through graded alcohol/water washes. Antigen retrieval was achieved by boiling samples 15min in citrate buffer (10 mM citric acid, 0.05% Tween 20, pH 6.0) prior to overnight primary antibody staining. Slides were mounted in Fluormount-G (Electron Microscopy Sciences). Slides were imaged on a Leica SP8 White light Laser Confocal microscope.

Balcioglu O., Gates B.L., Freeman D.W., Hagos B.M., Mehrabad E.M., Ayala-Talavera D, & Spike B.T. (2024). Mcam stabilizes luminal progenitor breast cancer phenotypes via Ck2 control and Src/Akt/Stat3 attenuation. bioRxiv.

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Quantifying β(1-3)-glucan in Candida cells

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Wild-type and cho1ΔΔ cells were grown in YPD overnight (~16 h), and back diluted to fresh YPD with OD₆₀₀ of ~0.1. The cells were then shaken at 225 rpm for 3 h before 170 µM CBR-5884 or equivalent DMSO were added. The cells were further shaken for 30, 60, 120 min before staining with anti-β(1-3)-glucan antibody. The cells were stained as previously described (57 (link), 97 (link)) with the exception that goat anti-mouse antibody conjugated to Alexa Fluor 488 (Jackson ImmunoResearch) was used as the secondary antibody. For imaging, Candida cells were resuspended in 200 µL of Fluoromount-G mounting medium and visualized with a Leica SP8 white light laser confocal microscope. The pictures were taken through Leica Application Suite X office software and quantified as described above.

Zhou Y., Phelps G.A., Mangrum M.M., McLeish J., Phillips E.K., Lou J., Ancajas C.F., Rybak J.M., Oelkers P.M., Lee R.E., Best M.D, & Reynolds T.B. (2024). The small molecule CBR-5884 inhibits the Candida albicans phosphatidylserine synthase. mBio, 15(5), e00633-24.

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Visualizing EGFR Signaling in A431 Cells

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A431 cells were plated at ∼80% confluency on a #1.5 glass coverslip, allowed to adhere for 24 h, and then starved overnight. Cells were then treated with serum-free DMEM without (no treatment) or with (PET1) PET1-DL680 for 1 h prior to a 5 min EGF 100 ng/ml treatment. Cells were washed with PBS containing 1 mM MgCl₂ and 100 mM CaCl₂ (PBS⁺⁺), fixed at 37 °C for 15 min in 4% paraformaldehyde, and permeabilized for 10 min at room temperature with 1% Triton X-100. Cells were blocked with 3% bovine serum albumin, and primary anti-EGFR XP (1:100) antibody was incubated overnight in 1% bovine serum albumin at 4 °C. Cells were washed, and secondary anti-rabbit conjugated to Alexa-fluor 488 (1:1000) was incubated 1 h at room temperature. Cells were then stained for DAPI (1 μg/ml) for 5 min and mounted to a microscope slide using Diamond Anti-fade mounting media. After curing, cells were imaged using a 63× 1.4NA oil objective on a Leica SP8 White Light Laser Confocal Microscope. Images were the product of 3-fold line averaging. Three to five images were taken per coverslip, and Pearson’s Correlation Coefficient (r) was calculated via the Coloc2 plugin from ImageJ.

Rybak J.A., Sahoo A.R., Kim S., Pyron R.J., Pitts S.B., Guleryuz S., Smith A.W., Buck M, & Barrera F.N. (2023). Allosteric inhibition of the epidermal growth factor receptor through disruption of transmembrane interactions. The Journal of Biological Chemistry, 299(7), 104914.

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