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Axiophot d1 microscope

Manufactured by Zeiss
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The Axiophot D1 is a high-performance microscope designed for advanced research applications. It features a modular design, allowing for customization to meet specific user requirements. The Axiophot D1 provides consistent optical performance and reliable operation, making it a versatile tool for a wide range of microscopy techniques.

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Lab products found in correlation

Axiocam mrc5 camera, by Zeiss (12 mentions) Axiocam mrc5, by Zeiss (3 mentions) Vanillin, by Merck Group (1 mentions) Ruthenium red, by Merck Group (1 mentions) Eclipse ti s, by Nikon (1 mentions) Progres mf cool monochromatic camera, by Jenoptik (1 mentions) Model c dsd230, by Nikon (1 mentions) Technovit 7100, by Kulzer (1 mentions) Dfc280 camera, by Leica (1 mentions) Dsm 940, by Zeiss (1 mentions) Axiocam mr camera, by Zeiss (1 mentions) Axiophot microscope, by Zeiss (1 mentions)

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17 protocols using axiophot d1 microscope

1

Seed Mucilage and PA Accumulation Analysis

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In order to check the PA accumulation, fresh seeds from silique at 3 days after pollination have been collected. Slides were mounted with a solution made by 1% (w/v) vanillin (Sigma-Aldrich, St. Louis, MI, USA, https://www.sigmaaldrich.com, accessed on 18 May 2021) and 5 M HCl and incubated at room temperature for 5 m. Samples were observed using a Zeiss Axiophot D1 microscope.
To check the mucilage extrusion, freshly harvested seeds were incubated for 15 m with a solution of 0.01% (w/v) ruthenium red (Sigma-Aldrich, https://www.sigmaaldrich.com, accessed on 18 May 2021). Samples were then analyzed using a Zeiss Axiophot D1 microscope.
Di Marzo M., Babolin N., Viana V.E., de Oliveira A.C., Gugi B., Caporali E., Herrera-Ubaldo H., Martínez-Estrada E., Driouich A., de Folter S., Colombo L, & Ezquer I. (2022). The Genetic Control of SEEDSTICK and LEUNIG-HOMOLOG in Seed and Fruit Development: New Insights into Cell Wall Control. Plants, 11(22), 3146.
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2

Microscopic Observation of Pollinated Pistils

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Emasculated pistils were observed 2 days after pollination, kept in 50 mM sodium phosphate buffer (pH 7.5) or water, and dissected under a stereomicroscope (Model C-DSD230, Nikon) using hypodermic needles (0.4 3 20 mm; Braun). The opened carpels and the ovules that remained attached to the septum were maintained in mounting medium and covered with a coverslip. Pistils pollinated with the LAT52 prom :GFP-H3.3:mRFP pollen were observed using a Zeiss Axiophot D1 microscope (http://www.zeiss.com) equipped with differential interference contrast optics. Images were recorded using an Axiocam MRc5 camera (Zeiss) with Axiovision version 4.1.
For phenotypic characterization, ovules at different developmental stages were cleared and analyzed as described previously (Brambilla et al., 2007) . Samples were observed using a Zeiss Axiophot D1 microscope equipped with differential interference contrast optics. Images were captured on an Axiocam MRc5 camera (Zeiss) using the Axiovision program (version 4.1). Aniline blue analyses of 35s prom :JAGGER pistils were observed under an inverted microscope (Eclipse Ti-S; Nikon) and images were captured with a ProgRes MF cool monochromatic camera (Jenoptik, Jena, Germany), and processed with ProgRes CapturePro 2.8.8 software.
Pereira A.M., Nobre M.S., Pinto S.C., Lopes A.L., Costa M.L., Masiero S, & Coimbra S. (2016). "Love Is Strong, and You're so Sweet": JAGGER Is Essential for Persistent Synergid Degeneration and Polytubey Block in Arabidopsis thaliana. Molecular plant, 9(4).
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3

Ovule Development Microscopy Protocol

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Inflorescences were fixed overnight in ethanol:acetic acid 9:1 and rehydrated in an ethanol series. Pistils (stages 7 to 10) were dissected and mounted in chloral hydrate:glycerol:water (8:1:3, w/v/v) and immediately observed under a Zeiss Axiophot D1 microscope (Carl Zeiss MicroImaging) equipped with DIC optics and an Axiocam MRc5 camera (Zeiss) with Axiovision software (version 4.1). Ovules were counted manually. Placenta length was measured using the Axiovision software. For statistical analysis in Fig. 1 wild type and hda19-3 were confronted by Student's t-test (two-tailed distribution, homoscedastic). For experiments in Figs. 5 and 6 the statistical significance of differences in ovule number and placenta length was determined by a one-way ANOVA and a Bonferroni post hoc test for multiple comparisons (P < 0.05).
Manrique S., Cavalleri A., Guazzotti A., Villarino G.H., Simonini S., Bombarely A., Higashiyama T., Grossniklaus U., Mizzotti C., Pereira A.M., Coimbra S., Sankaranarayanan S., Onelli E., Masiero S., Franks R.G, & Colombo L. (2023). HISTONE DEACETYLASE19 Controls Ovule Number Determination and Transmitting Tract Differentiation. Plant Physiology, 194(4), 2117-2135.
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4

Morphological Analysis of Arabidopsis Integuments

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For the morphological analysis of integuments, Arabidopsis thaliana wild-type (ecotype Columbia) and stk mutant plants were fixed for Technovit 7100 embedding (Heraeus Kulzer) following the manufacturer's instructions. Sections of plant tissue (0.8 µm) were stained in 0.5% (w/v) toluidine blue O. Samples were observed using a Zeiss Axiophot D1 microscope (http://zeiss.com/) equipped with differential interface contrast (DIC) optics. Images were recorded with an Axiocam MRc5 camera (Zeiss) using the Axiovision program (version 4.1). The whole-mount vanillin assay for PA detection was performed as described previously [61] (link). Vanillin (vanilaldehyde) condenses specifically with PAs and flavan-3-ol precursors to yield a bright-red product under acidic conditions. Microscopic observations were performed as detailed above.
Mizzotti C., Ezquer I., Paolo D., Rueda-Romero P., Guerra R.F., Battaglia R., Rogachev I., Aharoni A., Kater M.M., Caporali E, & Colombo L. (2014). SEEDSTICK is a Master Regulator of Development and Metabolism in the Arabidopsis Seed Coat. PLoS Genetics, 10(12), e1004856.
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5

GUS Histochemical Detection in Plants

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For GUS histochemical detection, plant material was fixed in 90% acetone at -20°C for 1 h. Samples were then washed three times with NaPi buffer (NaH2PO4 50 mM, Na2HPO4 50 mM; pH 7.0) and stained overnight at 37°C with X-gluc solution [1 mM 5-bromo-4-chloro-3-indolyl-β-D-clucoronide, 2 mM K3/K4Fe(CN)6, 0,1% Triton (v/v), 10 mM EDTA, 50 mM NaPi pH 7.0]. 70% EtOH (v/v) was used as washing solution. Stained samples were then stored at 4°C and observed using a Zeiss Axiophot D1 microscope equipped with differential interference contrast (DIC) optics. Images were recorded with an Axiocam MRc5 camera (Zeiss) using the Axiovision program (v.4.1).
Ferrari R., Tadini L., Moratti F., Lehniger M.K., Costa A., Rossi F., Colombo M., Masiero S., Schmitz-Linneweber C, & Pesaresi P. (2017). CRP1 Protein: (dis)similarities between Arabidopsis thaliana and Zea mays. Frontiers in Plant Science, 8, 163.
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6

Arabidopsis Ovule Clearing and Imaging

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Flowers were fixed with 9:1 ethanol:acetic acid and cleared overnight using a clearing solution of chloral hydrate:glycerol:water (3:1:2; w/v/v). Ovules were dissected from premature pistils after clearing, mounted with a cover slip, and subsequently observed in a Zeiss Axiophot D1 microscope equipped with DIC optics. Images were captured on an Axiocam MRc5 camera (Zeiss) using the Axiovision program (version 4.1). Three different plants of Arabidopsis ecotype Columbia and of each mutant, SALK_027711 and SALK_128927, were analyzed.
Ferreira L.G., Dusi D.M., Irsigler A.S., Gomes A.C., Florentino L.H., Mendes M.A., Colombo L, & Carneiro V.T. (2023). Identification of IPT9 in Brachiaria brizantha (syn. Urochloa brizantha) and expression analyses during ovule development in sexual and apomictic plants. Molecular Biology Reports, 50(6), 4887-4897.
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7

Histochemical Staining of Arabidopsis Siliques

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Arabidopsis siliques at stage 17 (Roeder and Yanofsky, 2006 (link)) were fixed in ethanol-acetic fixative (9:1) for 10 min under vacuum and kept overnight at 4°C. Samples were washed with 70% ethanol (30 min) and they were cleared in chloral hydrate:glycerol:H2O (8 g:1 mL:2 mL; w/v/v) solution for 24 h. Samples were dehydrated in a series of five increasing ethanol concentrations. Finally, Arabidopsis fruits were stained with 2% phloroglucinol solution in 96% ethanol for 5 min and then 50% HCl was applied for 1 min. Dissected fruits were then analyzed by Zeiss Axiophot D1 microscope equipped with differential interface contrast (DIC) optics and images were recorded with an Axiocam MRc5 camera (Zeiss) using the Axiovision program.
Moschin S., Nigris S., Ezquer I., Masiero S., Cagnin S., Cortese E., Colombo L., Casadoro G, & Baldan B. (2021). Expression and Functional Analyses of Nymphaea caerulea MADS-Box Genes Contribute to Clarify the Complex Flower Patterning of Water Lilies. Frontiers in Plant Science, 12, 730270.
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8

Microscopic Analysis of Vanillin-Stained Plant Tissues

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The vanillin assay for PAs detection was performed as described previously [32 (link)]. Vanillin (vanilaldehyde- V1104, Sigma-Aldrich, St. Louis, MO, USA) condenses with PAs and flavan-3-ol precursors to give a bright-red product in acidic conditions. Microscopic observations were performed using a Zeiss Axiophot D1 microscope equipped with differential interface contrast (DIC) optics. Images were recorded with an Axiocam MRc5 camera (Zeiss) using the Axiovision program (version 4.1).
Ruthenium-red (0.01% w/v Sigma-Aldrich) staining and evaluation of mucilage extrusion on mature seeds was performed upon 60 min of water imbibition as previously described [33 (link)]. Seeds were photographed with a Leica stereo microscope equipped with Leica DFC280 camera and imaging software LAS AF 2.2.0 (Leica UK Ltd., Milton Keynes, UK).
Scanning electron microscopy of mature dry seeds was performed as previously described [34 (link)] by gold coating them using a sputter coater (SEMPREP2; Nanotech) followed by observation with a LEO 1430 scanning electron microscope (LEO Electron Microscopy).
Paolo D., Orozco-Arroyo G., Rotasperti L., Masiero S., Colombo L., de Folter S., Ambrose B.A., Caporali E., Ezquer I, & Mizzotti C. (2021). Genetic Interaction of SEEDSTICK, GORDITA and AUXIN RESPONSE FACTOR 2 during Seed Development. Genes, 12(8), 1189.
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9

Morphological Analysis of Arabidopsis Inflorescences

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For morphological analysis, wild-type (ecotype E165) and era inflorescences and seedlings were fixed as previously described41 (link). Sections (8 μm) were stained in 0.5% (w/v) toluidine blue O or hybridized with digoxigenin-labelled ERA and STM antisense probes (primers are reported in Supplementary Table S1). DIG-labelled RNA probes were prepared as previously described41 (link). The samples were observed using a Zeiss Axiophot D1 microscope equipped with differential interface contrast (DIC) optics. Images were recorded with an Axiocam MRc5 camera (Zeiss) using the Axiovision program (version 4.1). Scanning electron microscopy (SEM) was performed by producing replicas of flowers and developing inflorescences as described by Green and Linstead42 . The samples were observed using a Zeiss DSM 940 scanning electron microscope.
Mizzotti C., Galliani B.M., Dreni L., Sommer H., Bombarely A, & Masiero S. (2017). ERAMOSA controls lateral branching in snapdragon. Scientific Reports, 7, 41319.
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10

Microscopic Analysis of Reproductive Organs

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Morphological analyses of reproductive organs -dissected from mature flowers at anthesis of different transgenic lines and wild-type control -were carried out by using a Zeiss Axiophot D1 microscope outfitted with a Zeiss AxioCam MR camera. Samples for histological analysis and wholemount tissue clearing were prepared as previously described (Dreni et al., 2007; (link)Dreni et al., 2011) (link).
Longitudinal thin sections (5-μm-thick) of wild-type and dst mutant mature pistils, previously embedded in Technovit resin, were stained with 0.1% Toluidin blue and mounted with a coverslip.
Osnato M., Lacchini E., Pilatone A., Dreni L., Grioni A., Chiara M., Horner D., Pelaz S, & Kater M.M. (2021). Transcriptome analysis reveals rice MADS13 as an important repressor of the carpel development pathway in ovules. Journal of experimental botany, 72(2).
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