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M165 fc fluorescent microscope

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The Leica M165 FC is a fluorescent microscope designed for advanced imaging applications. It features high-performance optics and a versatile illumination system to capture detailed fluorescent images. The M165 FC is a reliable and efficient tool for researchers and laboratory professionals.

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Lab products found in correlation

Dig rna labeling kit, by Roche (1 mentions) Dfc450 c digital camera, by Leica (1 mentions) Pgfp c shlenti plasmid, by OriGene (1 mentions)

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M165 FC fluorescent M165 FC Fluorescent microscope

7 protocols using m165 fc fluorescent microscope

1

Expression Profiling of Endogenous Genes near Insertion Sites

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To identify the expression profile of the endogenous gene near the insertion site, whole-mount RNA in situ hybridization (WISH) of the nearest gene was performed as previously described [36 (link)]. Antisense RNA probes for dlx1a, dlx2a, wnt1, wnt10b, rps26 and ednraa were synthesized by using digoxigenin (Roche, Basel, Switzerland) as a label. Images were captured using the M165 FC fluorescent microscope (Leica, Solms, Germany).
To investigate the transcription of the identified Ens, the embryos, developed to 24 and 48 hpf (hours post fertilization), were used for reverse transcription (RT)-PCR and real-time quantitative-PCR (qPCR) (the primers for qPCR are listed in Table S2).
Shen D., Xue S., Chan S., Sang Y., Wang S., Wang Y., Chen C., Gao B., Mueller F, & Song C. (2018). Enhancer Trapping and Annotation in Zebrafish Mediated with Sleeping Beauty, piggyBac and Tol2 Transposons. Genes, 9(12), 630.
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2

Whole-Mount In Situ Hybridization of Zebrafish Embryos

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To identify the expression profile of the endogenous gene itgav near the insertion site, the whole-mount in situ hybridization (WISH) on zebrafish embryos was performed as previously described [12 (link)]. Antisense RNA probes for target genes were synthesized according to the DIG RNA Labeling Kit (Roche). Embryos required for different periods were collected and fixed with paraformaldehyde (PFA). After permeabilization and prehybridization, hybridization was performed by incubating embryos with antisense DIG-labeled RNA probes overnight at 70 ℃, then followed by labeling reaction and staining, and images were captured using the M165 FC fluorescent microscope (Leica, Solms, Germany).
Jia W., Guan Z., Shi S., Xiang K., Chen P., Tan F., Ullah N., Diaby M., Guo M., Song C, & Gao B. (2022). The Annotation of Zebrafish Enhancer Trap Lines Generated with PB Transposon. Current Issues in Molecular Biology, 44(6), 2614-2621.
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3

Pollen Germination and Tube Growth Assay

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Four pollination treatments were conducted as above. Five pistils from each treatment group were collected at 2, 4, 6, 8, 12, 24, 48, 72, 96, 144, 168, and 192 h after pollination and were treated with fixative (three parts ethanol and one part glacial acetic acid) for at least 24 h, softened in 8 mol L−1 NaOH at 25 °C for 10 min, washed briefly three times in water, and stained with 0.1% water-soluble aniline blue dissolved in 0.1 mol K3PO4 for 1 h [59 (link)]. The pistils were assessed for pollen germination and pollen tube growth using a LEICA M165 FC fluorescent microscope (LEICA, Buffalo Grove, IL, USA) with a LEICA DFC450 C digital camera (LEICA, Buffalo Grove, IL, USA).
Lu W., Bian X., Yang W., Cheng T., Wang J., Zhang Q, & Pan H. (2018). Transcriptomics Investigation into the Mechanisms of Self-Incompatibility between Pin and Thrum Morphs of Primula maximowiczii. International Journal of Molecular Sciences, 19(7), 1840.
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4

Viral Infection in C. elegans: Asynchronous and Synchronous Methods

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Animals were either infected for four days as asynchronous populations or for two days as synchronous populations. Infections of asynchronous populations were performed as in 5 (link). Briefly, two L4 hermaphrodites were distributed in each 50 mm plates and, on the next day, 20 µl of viral filtrate was spread on the plates. Animals were harvested (for viral load measurement) or observed under a Leica M165 FC fluorescent microscope (for scoring of the viral stress sensor) four days post-infection (4 dpi). This method was typically used for the characterization of the Ovid screen isolates. For the infection of synchronous populations, 200 animals at the larval stage L1 were deposited on each 50 mm plate. On the next day, L2 animals were infected with 20 µl of viral filtrate homogeneously spread on the plate. Plates were kept up-side-up for 24 hrs. Animals were harvested for viral load measurement at 2 dpi. This method was used to measure the viral load in cde-1 mutants, as indicated in the figure legends.
Le Pen J., Jiang H., Di Domenico T., Kneuss E., Kosałka J., Leung C., Morgan M., Much C., Rudolph K.L., Enright A.J., O’Carroll D., Wang D, & Miska E.A. (2018). Terminal uridylyltransferases target RNA viruses as part of the innate immune system. Nature structural & molecular biology, 25(9), 778-786.
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5

Establishing Transgenic Ae. aegypti Mosquitoes

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The posterior pole of freshly laid pre-blastoderm Ae. aegypti embryos (<2 h old) were microinjected44 (link) using a Nikon Eclipse TS100 microscope and air pump (Jun-Air). The injection mix consisted of a final concentration of 500 ng/μl donor plasmid and 300 ng/μl helper plasmid [PUb hyperactive piggyBac transposase (AGG1245)]38 in 1x injection buffer. Microinjection survivors (G0) were screened for marker gene expression at the 4th instar larval stage using the Leica M165FC fluorescent microscope and appropriate filter setting, those transiently expressing the marker gene were individualised and mated with 3 wild-type mosquitoes. To identify transgenic lines the resultant G1 generation from these crosses were likewise screened at the 4th instar. Lines were maintained by mating virgin transgenic females with an excess of wild type males, offspring were screened at 4th instar stage.
McNamara C.J., Ant T.H., Harvey-Samuel T., White-Cooper H., Martinez J., Alphey L, & Sinkins S.P. (2024). Transgenic expression of cif genes from Wolbachia strain wAlbB recapitulates cytoplasmic incompatibility in Aedes aegypti. Nature Communications, 15, 869.
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6

Viral Infection in C. elegans: Asynchronous and Synchronous Methods

Cited 1 time
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Animals were either infected for four days as asynchronous populations or for two days as synchronous populations. Infections of asynchronous populations were performed as in 5 (link). Briefly, two L4 hermaphrodites were distributed in each 50 mm plates and, on the next day, 20 µl of viral filtrate was spread on the plates. Animals were harvested (for viral load measurement) or observed under a Leica M165 FC fluorescent microscope (for scoring of the viral stress sensor) four days post-infection (4 dpi). This method was typically used for the characterization of the Ovid screen isolates. For the infection of synchronous populations, 200 animals at the larval stage L1 were deposited on each 50 mm plate. On the next day, L2 animals were infected with 20 µl of viral filtrate homogeneously spread on the plate. Plates were kept up-side-up for 24 hrs. Animals were harvested for viral load measurement at 2 dpi. This method was used to measure the viral load in cde-1 mutants, as indicated in the figure legends.
Le Pen J., Jiang H., Di Domenico T., Kneuss E., Kosałka J., Leung C., Morgan M., Much C., Rudolph K.L., Enright A.J., O’Carroll D., Wang D, & Miska E.A. (2018). Terminal uridylyltransferases target RNA viruses as part of the innate immune system. Nature structural & molecular biology, 25(9), 778-786.
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7

Tumor Induction and Visualization on Chick CAM

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ISK, and HEC1A cells were labelled with an EGFP-expressing lentivirus donated by Dr Sokratis Theocharatos (University of Liverpool, UK). RL95-2 cells were labelled with a lentivirus expressing tGFP (pGFP-C-shLenti plasmid) (Origene, MD, USA), and selected using puromycin antibiotic selection. ISK and HEC1A viral transfection and egg preparation were performed according to Carter et al. 2012 [20] . Cells were added to the CAM at embryonic day 7 (E7) [21] the resulting tumours were imaged at E14, using a standard Leica M165-FC fluorescent microscope, in situ and after excision.
(Fig. 5C) the resulting tumours were fixed in 10% (v/v) NBF and embedded in paraffin wax.
Parkes C., Kamal A., Valentijn A.J., Alnafakh R., Gross S.R., Barraclough R., Moss D., Kirwan J, & Hapangama D.K. (2018). Assessing Estrogen-Induced Proliferative Response in an Endometrial Cancer Cell Line Using a Universally Applicable Methodological Guide. International journal of gynecological cancer : official journal of the International Gynecological Cancer Society, 28(1).
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