S8ap0 microscope

Manufactured by Leica

The Leica S8AP0 microscope is a high-performance stereo microscope designed for a variety of laboratory applications. It features a built-in apochromatic optical system, providing superior image quality and true-to-life color reproduction. The microscope offers a wide zoom range and long working distance, making it suitable for a wide range of sample sizes and types.

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Lab products found in correlation

Eos 40d camera, by Zeiss (2 mentions) Vhx 5000 digital microscope, by Keyence (2 mentions) Seaplaque agar, by Lonza (2 mentions) Crystal violet, by Merck Group (2 mentions) Ax10 microscope, by Zeiss (1 mentions) Buffered formalin, by Thermo Fisher Scientific (1 mentions) Formalin, by Thermo Fisher Scientific (1 mentions) 6 well plate, by Corning (1 mentions) Synergy mx, by Agilent Technologies (1 mentions) Mts assay, by Promega (1 mentions) Dfc295, by Leica (1 mentions) Ec4 camera, by Leica (1 mentions) C18 column, by Agilent Technologies (1 mentions)

9 protocols using s8ap0 microscope

Cave Beetle Morphometric Measurement Protocol

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The beetle specimens were collected by using an aspirator inside the cave, and kept in 55% ethanol before study. Dissections and observations were made under a Leica S8AP0 microscope. Dissected genital pieces, including the median lobe and parameres of the aedeagus, were glued onto small transparent plastic plates and pinned under the specimen they belonged to. Habitus pictures were taken by means of a Keyence VHX-5000 digital microscope. Genital pictures were taken using a Canon EOS 40D camera connected to a Zeiss AX10 microscope, and then stacked and processed by means of Adobe Photoshop CS5 software. Distribution maps created using Mapinfo software.
The length of the body was measured from the apex of the right mandible (in open position) to the elytral apex; the width of the body was taken as the maximum width of the elytra.
Abbreviations of other measurements used in the text are as follows:
HLm

length of head including mandibles, from apex of right mandible to occipital suture

HLl

length of head excluding mandibles, from front of labrum to occipital suture

maximum width of head

PrL

length of prothorax, along the median line

PnL

length of pronotum, as above

PrW

maximum width of prothorax

PnW

maximum width of pronotum

PfW

width of pronotum at front

PbW

width of pronotum at base

length of elytra, from base of scutellum to elytral apex

maximum width of combined elytra

Tian M., Huang S., Wang X, & Tang M. (2016). Contributions to the knowledge of subterranean trechine beetles in southern China’s karsts: five new genera (Insecta, Coleoptera, Carabidae, Trechinae). ZooKeys.

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Coleoptera Genital Dissection and Imaging

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The material was collected by using an aspirator inside the cave Shenxian Dong, and kept in 55% ethanol before study, except one of each species was kept in 95% ethanol for molecular analysis if two or more specimens were available. Dissections and observations were made under a Leica S8AP0 microscope. Dissected male genital pieces, including the median lobe and parameres of the aedeagus, were preserved in Euparal Mounting Medium (BioQuip Products, Inc., CA, USA) onto small transparent plastic plates and pinned under the specimen. Habitus pictures were taken by means of a Keyence VHX-5000 digital microscope. Genital pictures were taken using a Canon EOS 40D camera connected to a Zeiss AX10 microscope. Female genitalia were dissected before the entire abdomen was removed and placed in cold 10% KOH for one day, then cleaned in lactic acid for one day, and stained in Chlorazol Black which was dissolved in 70% ethanol for thirty seconds. All pictures were processed using Adobe Photoshop CS5 computer software.
Measurements and terminology follow Tian et al. (2016) (link). Terminology for female reproductive tract follows Deuve (1993 , 2018 (link)) and Liebherr and Will (1998) .

Fang J., Li W., Wang X, & Tian M. (2020). New cavernicolous ground beetles from Anhui Province, China (Coleoptera, Carabidae, Trechini, Platynini). ZooKeys, 923, 33-50.

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Quantifying Anchorage-Independent Cell Growth

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A layer of sea plaque agar (Lonza, Basel) at a concentration of 0.8% in culture media was placed into 6-well plates and allowed to solidify. A top layer was then placed into each well containing 10,000 cells in 0.48% agar in culture media and allowed to solidify. One mL of culture media was placed on top of the solidified agar with or without 2.5 μg doxycycline/mL of media. Plates were incubated for 2 weeks, with one mL of culture media added to every well after one week. After two weeks, the media was removed and cells were fixed in 10% buffered formalin (Thermo Fisher) containing 0.5% crystal violet for 2 hours. Each well was divided into four quadrants and photographed on a Leica S8 AP0 microscope. Colonies were quantified using ImageJ software and graphed using the Prism software package. Three wells were used for each condition, conducted in triplicate, giving 9 wells per condition and 36 quadrants for imaging. Two-tailed, unpaired t-tests with α = 0.05 were used to identify statistically significant differences in colony numbers (p < 0.05).

Marko T.A., Shamsan G.A., Edwards E.N., Hazelton P.E., Rathe S.K., Cornax I., Overn P.R., Varshney J., Diessner B.J., Moriarity B.S., O’Sullivan M.G., Odde D.J, & Largaespada D.A. (2016). Slit-Robo GTPase-Activating Protein 2 as a metastasis suppressor in osteosarcoma. Scientific Reports, 6, 39059.

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Soft Agar Colony Formation Assay

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6-well plates (Corning) were prepared with bottom agar (3.2%) and top agar (0.48%) composed of low melting point agarose in DMEM full media. The bottom agar was allowed to solidify before 10,000 cells in top agar were plated and allowed to solidify. DMEM media supplemented with 10% FBS and 2 ug/ml puromycin was plated over the cells in the 6-well plates and incubated under standard conditions. After 14 days, top media were removed, and cells were fixed in 10% formalin (Fisher Scientific) containing 0.0005% crystal violet (Sigma) for 1 h at room temperature. Colonies were imaged on Leica S8 AP0 microscope with 12 images per cell line. Automated colony counts were done using ImageJ software^{66 (link)} using TKS Batch Count Colonies macro (courtesy of StarrLab). Results shown are a representative example of at least 3 independent experiments.

Suppiah S., Mansouri S., Mamatjan Y., Liu J.C., Bhunia M.M., Patil V., Rath P., Mehani B., Heir P., Bunda S., Velez-Reyes G.L., Singh O., Ijad N., Pirouzmand N., Dalcourt T., Meng Y., Karimi S., Wei Q., Nassiri F., Pugh T.J., Bader G.D., Aldape K.D., Largaespada D.A, & Zadeh G. (2023). Multiplatform molecular profiling uncovers two subgroups of malignant peripheral nerve sheath tumors with distinct therapeutic vulnerabilities. Nature Communications, 14, 2696.

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Proliferation and Colony Formation Assays

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Proliferation assays were set up in a 96-well format with 100 cells plated per well. Proliferation was assessed every 24 hours over 6 days by the MTS assay, following manufactures instructions (Promega). Absorbance was read at 490 nm to determine proliferation and 650 nm to account for cellular debris on a BioTek Synergy Mx automated plate reader. Soft agar assays were performed as follows, 6-well plates were prepared with bottom agar composed of 3.2% SeaPlaque Agar (Lonza) in DMEM full media and allowed to solidify before 10,000 cells in top agar (0.8% SeaPlaque Agar in DMEM full media) were plated and allowed to solidify. DMEM full media with 2.5 µg/mL doxycycline was plated over the cells and cells were incubated under standard conditions (5% CO₂,37°C) for 2 weeks. Top media was removed and cells were fixed in 10% formalin (Fisher Scientific) containing 0.005% crystal violet (Sigma) for 1 hour at room temperature. Formalin was removed and colonies were imaged on a Leica S8 AP0 microscope. 12 images per cell line were taken and automated colony counts were done using ImageJ software. Results shown are a representative example of at least 3 independent experiments.

Moriarity B.S., Rahrmann E.P., Beckmann D.A., Conboy C.B., Watson A.L., Carlson D.F., Olson E.R., Hyland K.A., Fahrenkrug S.C., McIvor R.S, & Largaespada D.A. (2014). Simple and Efficient Methods for Enrichment and Isolation of Endonuclease Modified Cells. PLoS ONE, 9(5), e96114.

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Discovering a New Microlocia Species in Colombia

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This new species was found in the course of reviewing collections in regional herbaria of Colombia generated by the Boyacá BIO project. After several collections were located in the Federico Medem (FMB, Villa de Leyva) and Universidad Distrital (UDBC, Bogotá) herbaria, it was possible to visit the localities and collect new samples. Therefore new collections, field photographs and samples of flowers, fruits, and seeds stored in 70% alcohol were made.
Measurements of vegetative parts were made in the dry herbarium material with a digital caliper with a precision of 0.1 mm. The measurements of floral parts were based on fresh flowers preserved in alcohol from the type specimens. A Leica S8AP0 microscope was used. Photographs of leaves, flowers, fruits, and seeds were taken in the field and laboratory from fresh material using an MC190 HD camera.
The distribution of the Microlocia was associated with South America wind currents to evaluate the possible cause of the presence of the new taxon in the Andes of Colombia. For this, the map of winds patterns from the Earth Nullschool (https://earth.nullschool.net) was used, on which the distribution records of Microlicia from the electronic Tropicos database of the Missouri Botanical Garden (http://www.tropicos.org) were mapped using Arc-GIS version 10.2.1.

Mendoza-Cifuentes H., Ariza W., Granados D.E, & Rosana R.o.m.e.r.o. (2019). A new species of Microlicia (Melastomataceae): first record of the genus for Colombia. PhytoKeys, 122, 87-96.

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Visualizing Auxin Signaling in Transgenic Plants

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Transgenic MT and CKX2-OE double transgenic DR5::GUS plants were incubated overnight at 37 °C in GUS staining solution [80 mM sodium phosphate buffer, pH 7.0; 0.4 mM potassium ferrocyanide; 8 mM EDTA; 0.05% Triton X-100; 0.8 mg/mL 5-bromo-4-chloro-3-indolyl-b-D-glucuronide (X-Gluc); 20% methanol]. After GUS staining, samples were washed in 70% (w/v) ethanol to remove chlorophyll. Samples were then photographed using a Leica S8AP0 microscope coupled to a Leica DFC295 camera.

Pino L.E., Lima J.E., Vicente M.H., de Sá A.F., Pérez-Alfocea F., Albacete A., Costa J.L., Werner T., Schmülling T., Freschi L., Figueira A., Zsögön A, & Peres L.E. (2022). Increased branching independent of strigolactone in cytokinin oxidase 2-overexpressing tomato is mediated by reduced auxin transport. Molecular Horticulture, 2, 12.

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Early Embryo Development Assessment

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To assess the effects of QS10, idebenone and rotenone during early stages of embryo development, the number of spontaneous coiling events observed in 15 s were recorded at 28 hpf using the Leica S8AP0 microscope. To evaluate survival rate, 75 embryos from each group were observed every 10 hours at 20, 30, 40, 50, 60, 70, 80 hpf. The number of surviving embryos was counted at each time point.

Giorgio V., Schiavone M., Galber C., Carini M., Da Ros T., Petronilli V., Argenton F., Carelli V., Acosta Lopez M.J., Salviati L., Prato M, & Bernardi P. (2018). The idebenone metabolite QS10 restores electron transfer in complex I and coenzyme Q defects. Biochimica et biophysica acta. Bioenergetics, 1859(9).

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Phenotypic Responses to Predator Cues

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Experimental exposures: Phenotypic data were collected from 105 iso-female lines from the Killwood Nature Preserve (Suppl. Tab. 2). To establish predation risk conditions, we generated predator cues from frozen midge larvae following established protocols [62] . Homogenized midge larvae extracts were filtered, followed by solid-phase extraction using a C18 column (Agilent) to recover the active compounds that generate strong morphological responses in D. pulex. For experimental exposures, animals were kept under standard conditions for three generations. Subsequently, at least two gravid Daphnia, carrying embryos in E3 stage (~18 hours before parturition; sensu [63] ), were placed in individual jars containing 50 ml hard artificial pond water [52] , algae (2x10 5 cells ml -1 Chlorella vulgaris), liquid seaweed extract and 0 or 0.5 µl ml -1 Chaoborus predator cue concentrate. After parturition, two neonates were randomly selected from each of the two mothers per treatment (Suppl. Fig. 2) and placed individually in 50 ml glass vials containing the same medium as their maternal environment. For three to four consecutive days, each animal was photographed daily (Leica S8AP0 microscope; Leica EC4 camera) and subsequently transferred to a new glass vial containing fresh media and predator cues.

Becker D., Barnard-Kubow K., Porter R., Edwards A., Voss E., Beckerman A.P., & Bergland A.O. (2021). Stabilizing selection shapes variation in phenotypic plasticity.

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