The backscattered scanning electron microscopy (bSEM) procedures were described previously (Smith et al. 2011c ). For enamel thickness measurements, soft tissues were removed from left and right hemimandibles of wild‐type, Klk4−/−, Mmp20−/− and Mmp20−/−Klk4−/− (MK double null) mice at 7 weeks, sectioned at the level of the labial alveolar ridge (Fig. S2), and imaged by bSEM. Incisor and molar imaging was performed at 7 weeks on wild‐type, Klk4+/−, Klk4−/−, Mmp20+/−, Mmp20−/−, and Mmp20−/−Klk4−/− (MK double null) mice. For incisor imaging, the bony caps and soft tissue covering the mandibular incisors were carefully removed, and examined at ×50 magnification in a Hitachi S‐3000N variable pressure scanning electron microscope using the backscatter mode at 25 kV and 20 pascal pressure. For molar imaging, soft tissues were dissected away, the crowns wiped clean and air‐dried and then imaged in backscatter mode at ×40 magnification using 15–30 kV and 20 pascal pressure.
S 3000n variable pressure scanning electron microscope
Manufactured by Hitachi
Sourced in Japan
The S-3000N is a variable pressure scanning electron microscope manufactured by Hitachi. It is designed to observe and analyze a wide range of samples, including non-conductive materials, without the need for extensive sample preparation. The S-3000N allows for adjustable vacuum pressure, enabling the examination of samples in their natural state.
Automatically generated - may contain errors
Lab products found in correlation
Model hcp 2, by Hitachi (2 mentions)
Smz1000 stereoscopic microscope, by Olympus (1 mentions)
Bx60 microscope, by Nikon (1 mentions)
A1 confocal microscope, by Nikon (1 mentions)
Ds ri1 camera, by Nikon (1 mentions)
E 1010 ion sputter coater, by Hitachi (1 mentions)
Double sided carbon tape, by Agar Scientific (1 mentions)
8 protocols using s 3000n variable pressure scanning electron microscope
1
Backscattered Scanning Electron Microscopy Procedures
2
Scanning Electron Microscopy of P10 Brains
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Brains collected at P10 were fixed in Karnovsky’s fixative (2.5% glutaraldehyde, 2% paraformaldehyde, 0.1 M sodium cacodylate buffer pH 7.4) for 7 days at 4°C. Brain ventricles were then split and fixed in Karnovsky’s for 4 additional days. Brains were washed in 0.1 M cacodylate buffer and dehydrated through series of ethanol washes. Dehydrated tissue was immersed in pure hexamethyldisilazane (HMDS) (EMS, Hatfield, PA) twice for 15 minutes at RT. Samples were covered in fresh HMDS and left under the hood until evaporated. Specimens were mounted and sputter coated with gold/palladium using a Polaron E5100 coater (Polaron Instruments, West Sussex, UK). Images were taken using a Hitachi S-3000N variable pressure scanning electron microscope (Hitachi, Tokyo, Japan).
3
Confocal and Electron Microscopy Imaging
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Confocal microscopy images were taken with a Nikon A1 confocal microscope. Samples were either live-imaged or fixed and sectioned as previously described [9 (link)]. Excitation and detection wavelengths for GFP, Venus, and DsRed were as previously described [9 (link), 32 (link)]. To detect FM4-64 and PI staining, a 514 nm laser line was used for excitation and a 561 nm long-pass filter was used for detection. The modified pseudo-Schiff-PI (mPS-PI) staining was performed as described and a 488 nm laser line was used for excitation and emission was collected at 520–720 nm [62 (link)]. DAPI staining was excited at 405 nm and detected in the 425–475 nm. Autofluorescence was excited at 488 nm or 514 nm and detected in the 660–700 nm range.
Optical photographs were taken with a Nikon SMZ1000 stereoscopic microscope or an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera. Scanning electron microscopy was performed using a Hitachi S-3000N variable pressure scanning electron microscope after standard tissue preparation [9 (link)].
Optical photographs were taken with a Nikon SMZ1000 stereoscopic microscope or an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera. Scanning electron microscopy was performed using a Hitachi S-3000N variable pressure scanning electron microscope after standard tissue preparation [9 (link)].
4
Petal Sample Preparation for SEM Imaging
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Petal samples were collected by cutting into small pieces and fixed in FAA solution (formalin: glacial acetic acid: 70% ethanol = 1:1:18) as described [41 (link)]. The fixed samples were dehydrated by going through the gradual ethanol series, and then dried by critical point drying method by liquid carbon dioxide (Model HCP-2, Hitachi, Japan) and then gold-coated by an Edwards E-1010 ion sputter coater (Hitachi, Japan). The samples were observed with a S-3000 N variable pressure scanning electron microscope (Hitachi, Japan).
5
Scanning Electron Microscopy Sample Preparation
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Samples were fixed in 2.5% glutaraldehyde solution. Fixed samples were dehydrated with gradual ethanol series, dried by critical-point drying method using liquid carbon dioxide (Model HCP-2, Hitachi, Tokyo), gold-coated with an Edwards E-1010 ion sputter coater (Hitachi, Tokyo), and then observed using a S-3000 N variable pressure scanning electron microscope (Hitachi, Tokyo).
6
Backscattered SEM Imaging of Mouse Incisors and Molars
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This backscattered scanning electron microscopy (bSEM) procedures were described previously (Smith et al. 2011a (link)). Soft tissue was removed from the left and right hemimandibles of Wdr72+/+, Wdr72+/−, and Wdr72−/− mice at 7 weeks, and cross-sectioned at 1 mm increments from the basal (growing) end of the incisors, and imaged by bSEM. For whole incisor surface imaging, the bony caps and soft tissue covering the mandibular incisors were carefully removed, and examined at 50× magnification with a Hitachi S-3000N variable pressure scanning electron microscope using the backscatter mode at 25 kV and 20 pascal pressure.
Wdr72+/+, Wdr72+/−, and Wdr72−/− mouse molars were prepared as follows: The D14 mandibles were submerged in 4% PFA overnight; 7-week-old mice were fixative perfused with 4% PFA. The hemimandibles were carefully dissected of soft tissues under a stereoscopic microscope using tissue forceps and a spoon excavator, submerged in 1% NaClO for 5 min, rinsed, and acetone dehydrated (30%, 50%, 70%, 80%, 90%, 100%). The hemimandibles were mounted on metallic stubs using conductive carbon cement and carbon coated to increase conductivity and examined using a Hitachi (Century City, Los Angeles, CA) S-3000N variable pressure scanning electron microscope using the backscatter mode.
Wdr72+/+, Wdr72+/−, and Wdr72−/− mouse molars were prepared as follows: The D14 mandibles were submerged in 4% PFA overnight; 7-week-old mice were fixative perfused with 4% PFA. The hemimandibles were carefully dissected of soft tissues under a stereoscopic microscope using tissue forceps and a spoon excavator, submerged in 1% NaClO for 5 min, rinsed, and acetone dehydrated (30%, 50%, 70%, 80%, 90%, 100%). The hemimandibles were mounted on metallic stubs using conductive carbon cement and carbon coated to increase conductivity and examined using a Hitachi (Century City, Los Angeles, CA) S-3000N variable pressure scanning electron microscope using the backscatter mode.
7
Scanning Electron Microscopy of Tomato Floral Organs
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Floral organs except for gynoecia were removed from wild-type and mutant tomato flowers, and the samples were fixed in 100% methanol for 15 min and dehydrated in 100% ethanol for 30 min at room temperature. After one change of 100% ethanol, the tissues were stored in 100% ethanol overnight at room temperature. Tissues were dried with CO 2 in a critical point drier and coated with gold in a sputter coater. Tissues were imaged using a Hitachi S-3000N variable pressure scanning electron microscope at an accelerating voltage of 5 kV.
8
SEM Analysis of Spray-Dried and Freeze-Dried Proteins
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The morphologies of the spray-dried and freeze-dried protein particles were inspected using scanning electron microscope (SEM) (Hitachi S3000-N variable pressure scanning electron microscope, Japan). Small quantity of the dried protein preparations were attached to a double-sided carbon tape (Agar Scientific, Stansted, UK), positioned on an aluminium stub.
The samples were coated with a mixture of gold/palladium using a Quorum Technology (Polaron Range) SC760 by exposing samples to an Argon atmosphere at about 10 -1 mbar or 10 Pa. Samples were coated for 2 × 105 s.
The samples were coated with a mixture of gold/palladium using a Quorum Technology (Polaron Range) SC760 by exposing samples to an Argon atmosphere at about 10 -1 mbar or 10 Pa. Samples were coated for 2 × 105 s.
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