Pen slides

Manufactured by Leica camera

The PEN slides are a series of photographic equipment designed for professional and advanced amateur photographers. The PEN slides provide a convenient and efficient way to view and store photographic slides. The slides are made of high-quality materials and are designed to protect the slides from damage during storage and handling.

Automatically generated - may contain errors

Lab products found in correlation

Nanozoomer s60 slide scanner, by Hamamatsu Photonics (3 mentions) View2, by Hamamatsu Photonics (2 mentions) Paxgene tissue fix, by Qiagen (2 mentions) Lmd6500, by Leica camera (1 mentions) Rnalater, by Merck Group (1 mentions) High pure ffpet rna isolation kit, by Roche (1 mentions) Hematoxylin staining, by Vector Laboratories (1 mentions) Eosin y, by Thermo Fisher Scientific (1 mentions) As lmd laser capture microscope, by Leica camera (1 mentions) Lcm staining kit, by Thermo Fisher Scientific (1 mentions) Nextera xt kit, by Illumina (1 mentions) Rnalater, by Thermo Fisher Scientific (1 mentions) Hiseq 2500, by Illumina (1 mentions) Ndp view2, by Hamamatsu Photonics (1 mentions)

9 protocols using pen slides

Microdissection and Analysis of Plasmablast Patches

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7 µm spleen sections were prepared from OCT-frozen tissues on the membrane-coated PEN slides (Leica). Plasmablast patches were detected using anti-CD138 and GC-like structures using PNA and anti-IgD in immunohistochemistry as reported in Supplemental Information. Microdissections were performed using a Leica LMD6500 instrument equipped with an optical microscope. Dissected patches were collected in the cap of PCR microtubes in 10 ml of digestion buffer (50 mM Tris-HCl, 50 mM KCl, 0.63 mM EDTA, 0.22% Igepal, 0.22% Tween20, 0.8 mg/ml proteinase K). Patches were digested at 55°C for 2 hr, then at 90°C for 5 min, and used for PCR amplification of Ab genes. Primers, PCR procedures, and data analysis are described in Supplemental Information.

Di Niro R., Lee S.J., Vander Heiden J.A., Elsner R.A., Trivedi N., Bannock J.M., Gupta N.T., Kleinstein S.H., Vigneault F., Gilbert T.J., Meffre E., McSorley S.J, & Shlomchik M.J. (2015). Salmonella Infection Drives Promiscuous B Cell Activation Followed by Extrafollicular Affinity Maturation. Immunity, 43(1), 120-131.

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Histological Tissue Processing and Staining

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Tissues were embedded in Optimal Cutting Temperature compound, and frozen histological sections were cut at 30 µm, mounted on polyethylene naphthalate (PEN) slides and fixed in 70% ethanol for 5 min, followed by two washes with PBS for 1 min each. Slides were manually stained in hematoxylin and eosin (H&E) using a conventional staining protocol. A subset of samples (PD44594c–h and PD44589f) were fixed in PAXgene Tissue FIX (Qiagen) according to the manufacturer’s instructions. Fixed tissue samples were embedded in paraffin using a Tissue-Tek tissue-processing machine (Sakura). No formalin was used in the preparation, storage, fixation or processing of samples. Processed tissue blocks were embedded in paraffin wax, sectioned to 10-µm thickness and mounted onto PEN slides (Leica). Tissue slides were stained using a standard H&E protocol. Slides were temporarily coverslipped and scanned on a NanoZoomer S60 Slide Scanner (Hamamatsu); images were viewed using NDP.View2 software (Hamamatsu).

Robinson P.S., Coorens T.H., Palles C., Mitchell E., Abascal F., Olafsson S., Lee B.C., Lawson A.R., Lee-Six H., Moore L., Sanders M.A., Hewinson J., Martin L., Pinna C.M., Galavotti S., Rahbari R., Campbell P.J., Martincorena I., Tomlinson I, & Stratton M.R. (2021). Increased somatic mutation burdens in normal human cells due to defective DNA polymerases. Nature Genetics, 53(10), 1434-1442.

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Tissue Preparation and Staining Protocol

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Tissues were embedded in Optimal Cutting Temperature (OCT) compound, frozen histological sections were cut at 25–30 µm and mounted on polyethylene naphthalate (PEN) slides and fixed in 70% ethanol for 5 minutes followed by two washes with phosphate buffered saline for 1 min each. Slides were manually stained in haematoxylin and eosin using a conventional staining protocol. A subset of samples were fixed in RNAlater (Sigma Aldrich) according to manufacturer’s instructions. Fixed tissue samples were embedded in paraffin using a Tissue-Tek tissue processing machine (Sakura). No formalin was used in the preparation, storage, fixation or processing of samples. Processed tissue blocks were embedded in paraffin wax, sectioned to 10 µm thickness and mounted onto PEN slides (Leica). Tissue slides were stained using a standard haematoxylin and eosin (H&E) protocol. Slides were temporarily cover-slipped and scanned on a NanoZoomer S60 Slide Scanner (Hamamatsu), images were viewed with NDP.View2 software (Hamamatsu).

Robinson P.S., Thomas L.E., Abascal F., Jung H., Harvey L.M., West H.D., Olafsson S., Lee B.C., Coorens T.H., Lee-Six H., Butlin L., Lander N., Truscott R., Sanders M.A., Lensing S.V., Buczacki S.J., ten Hoopen R., Coleman N., Brunton-Sim R., Rushbrook S., Saeb-Parsy K., Lalloo F., Campbell P.J., Martincorena I., Sampson J.R, & Stratton M.R. (2022). Inherited MUTYH mutations cause elevated somatic mutation rates and distinctive mutational signatures in normal human cells. Nature Communications, 13, 3949.

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Profiling UVB-Induced Transcriptional Changes

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Six millimetre punch biopsies were taken from psoriatic plaques or adjacent non-lesional skin at 6 h (± 2 h) or 18 h (± 1 h) after irradiation with 311 nm or 290 nm UVB and from unirradiated psoriatic plaque skin. A maximum of four biopsies were taken per patient. Biopsies were snap frozen, set in OCT embedding medium (Raymond A Lamb) and stored at -80 °C. 35 μm cryosections were cut, collected on PEN slides (Leica, Milton Keynes) and kept frozen until microdissection. Slides were fixed in ethanol and stained with toluidine blue, and then the epidermis was micro-dissected with a scalpel using a dissection microscope. RNA was extracted using a Picopure TM RNA Isolation kit (Arcturus Biosciences, USA) using the manufacturers standard protocol, and stored at -80 °C.

Addison R., Weatherhead S.C., Pawitri A., Smith G.R., Rider A., Grantham H.J., Cockell S.J, & Reynolds N.J. (2021). Therapeutic wavelengths of ultraviolet B radiation activate apoptotic, circadian rhythm, redox signalling and key canonical pathways in psoriatic epidermis. Redox Biology, 41, 101924.

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Laser Capture Microdissection of Cerebellum

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10 µm-thick sections for normal (n=2) and DWM (n=2) cerebella embedded in paraffin were collected on 2.0 µm PEN slides (Leica No.11505189). These were subsequently stained with 1% Cresyl Violet after deparaffinization with Xylene and washes in Ethanol. Laser capture microdissection was performed as described previously [18 (link)]. The microdissected tissue was transferred into RNA tissue lysis buffer provided in the High Pure FFPET RNA Isolation Kit (Roche , ref.06650775001) and RNA extraction performed using manufacturer-recommended protocols. The Agilent Bioanalyzer 600 Pico Kit was used to assess RNA quality (DV200 > 74 for all samples).

Haldipur P., Bernardo S., Aldinger K.A., Sivakumar T., Millman J., Sjoboen A.H., Dang D., Dubocanin D., Deng M., Timms A.E., Davis B.D., Plummer J.T., Mankad K., Oztekin O., Manganaro L., Guimiot F., Adle-Biassette H., Russo R., Siebert J.R., Kidron D., Petrilli G., Roux N., Razavi F., Glass I.A., Di Gioia C., Silvestri E, & Millen K.J. (2021). Evidence of disrupted rhombic lip development in the pathogenesis of Dandy-Walker malformation. Acta neuropathologica, 142(4), 761-776.

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FFPE Tissue Preparation for Laser Capture

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Formalin-fixed paraffin-embedded (FFPE) tissue blocks were sectioned at (10 µm thick) and mounted onto Leica PEN slides (Cat No. 11505189). Slides were deparaffinized using 100% xylene (2x), 100% ethanol, 95% ethanol, 70% ethanol and 50% ethanol (3 min each). This preparation was followed by hematoxylin staining (Vector) for 1 min. Slides were then rinsed in de-ionized water (1 min) and stained in 1% eosin Y (Fisher scientific) and 1% calcium chloride (6 min). Slides were then left to air dry for laser capture microdissection.

Lam K.H., Leon A.J., Hui W., Lee S.C., Batruch I., Faust K., Klekner A., Hutóczki G., Koritzinsky M., Richer M., Djuric U, & Diamandis P. (2022). Topographic mapping of the glioblastoma proteome reveals a triple-axis model of intra-tumoral heterogeneity. Nature Communications, 13, 116.

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Laser Capture Microscopy of Human Epidermis

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Clinically normal human epidermis of sun-protected truncal sites was collected from elective surgeries. Duplicate samples from unrelated adults were placed into RNALater (Ambion) and embedded into O.C.T. (Sakura), and 7-µM sections were mounted on PEN slides (Leica). Slides were stained using an LCM staining kit (Ambion), and sections were harvested with a Leica AS LMD laser capture microscope. Samples were processed with the SMART-Seq version 3 ultralow-input RNA kit (Clontech), and sequencing libraries were prepared with the Nextera XT kit (Illumina). Sequencing was performed on an Illumina HiSeq 2500 with 50-base-pair single-end reads. Sequence data have been deposited at National Institutes of Health Sequence Read Archive (SRA 064531, 064538, and 064547). Please see Supplemental Figure 6 for discussion of RNA-seq analysis methods.

Sun B.K., Boxer L.D., Ransohoff J.D., Siprashvili Z., Qu K., Lopez-Pajares V., Hollmig S.T, & Khavari P.A. (2015). CALML5 is a ZNF750- and TINCR-induced protein that binds stratifin to regulate epidermal differentiation. Genes & Development, 29(21), 2225-2230.

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Single-Cell Laser Microdissection of GFP-Labeled Neurons

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A Leica Laser Microdissection system (LMD7000) was used to isolate single GFP-labeled PNs. We re-identified single target PNs that had been previously analyzed in 60 μm-thick slices in the re-sectioned 25 μm-thick sections, based on their relative spatial position and dendritic geometry. PEN slides (Leica) were loaded onto the microscope with a 63 × objective (HCX PL FLUOTAR NA: 0.7, Leica) and contours of target single cells were hand-drawn onto the slice image displayed on the computer monitor using a tablet and a stylus. The laser was directed along the border of single cell, and the isolated cell was collected into a lid of 0.5 mL tubes. Laser parameters were adjusted as follows: power 40, aperture 1, speed 1, specimen balance 25, head current 80%, pulse frequency 1019, and offset 210.

Steinecke A., Kurabayashi N., Hayano Y., Ishino Y, & Taniguchi H. (2019). In Vivo Single-Cell Genotyping of Mouse Cortical Neurons Transfected with CRISPR/Cas9. Cell reports, 28(2), 325-331.e4.

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Histological Tissue Preparation and Staining

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Tissues were embedded in Optimal Cutting Temperature (OCT) compound, frozen histological sections were cut at 30µm and mounted on polyethylene naphthalate (PEN) slides and fixed in 70% ethanol for 5 minutes followed by two washes with phosphate buffered saline for 1 minute each. Slides were manually stained in haematoxylin and eosin using a conventional staining protocol. A subset of samples (PD44594c-h and PD44589f) were fixed in PAXgene Tissue FIX (Qiagen) according to manufacturer's instructions. Fixed tissue samples were embedded in paraffin using a Tissue-Tek tissue processing machine (Sakura). No formalin was used in the preparation, storage, fixation or processing of samples. Processed tissue blocks were embedded in paraffin wax, sectioned to 10µm thickness and mounted onto PEN slides (Leica). Tissue slides were stained using a standard haematoxylin and eosin (H&E) protocol. Slides were temporarily cover-slipped and scanned on a NanoZoomer S60 Slide Scanner (Hamamatsu), images were viewed with NDP.View2 software (Hamamatsu).

Robinson P.S., Coorens T., Palles C., Mitchell E., Abascal F., Ólafsson S., Lee B.T., Lawson A., Lee-Six H., Moore L., Sanders M.A., Hewinson J., Martin L., Pinna C.M., Galvotti S., Campbell P.J., Martincorena I., Tomlinson I., & Stratton M.R. (2020). Elevated somatic mutation burdens in normal human cells due to defective DNA polymerases.

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