The excised radical prostatectomy specimens were fixed in formalin for 48 h and then sectioned and processed. Each prostate was hand sectioned into 2–3 mm thick axial slices, embedded in paraffin, and a microtome was used to cut a 5 λm thin single slice from each thick section. The thin slices were then attached to a glass slide that was processed with Hematoxylin and Eosin staining. A pathologist who was blinded to all imaging data, identified and marked the left–right and anterior–posterior aspect of each slide as well as anatomical structures including the urethra and verumontanum. He also identified BPH, atrophy, and PCa. Each prostate specimen yielded 8–21 whole mount slides. The histology slides were then digitized (0.04 × 0.04 mm) using a flat-bed scanner (Epson Perfection V750-M Pro Scanner with 640 dpi resolution).
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6 protocols using perfection v750 m pro scanner
1
Histological Analysis of Prostate Cancer
2
Cultivation of C. glutamicum in CGXII
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C. glutamicum was precultured in BHI or LB medium overnight, washed once with CGXII medium (Eggeling and Bott, 2005 (link)) without carbon source and inoculated in CGXII with 222 mM of glucose at initial OD at 600 nm of 1. The OD was measured with UV-1202 spectrophotometer (Shimadzu, Duisburg, Germany) with suitable dilutions. When appropriate, 100 μg/mL of spectinomycin, 25 μg/mL of kanamycin and IPTG were added. Growth experiment with Biolector® cultivation system (m2pLabs, Baesweiler, Germany) was performed in 1 mL of CGXII using FlowerPlate® (m2pLabs, Baesweiler, Germany) at 30°C, 1,100 rpm. Cell growth was monitored online every 10 min for 48 h. Maximum growth rate μ (h-1) was calculated from 20 measuring points of arbitrary unit of backscattering light (620 nm). Plate image was scanned with Perfection V750-M Pro scanner (Epson, Ludwigshafen am Rhein, Germany). Color balance of blue against yellow was set to +70.
3
Growth Kinetics of C. glutamicum
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As far as not mentioned specifically, C. glutamicum was precultured in LB medium (Sambrook, 2001 ) with 56 mM of glucose overnight, washed once with CGXII medium (Eggeling and Bott, 2005 ) without carbon source and inoculated in CGXII with 222 mM of glucose at initial optical density (OD) (λ=600 nm) of 1. The OD was measured with UV-1202 spectrophotometer (Shimadzu, Duisburg, Germany) with suitable dilutions. When appropriate, 25 μg/mL of kanamycin and IPTG were added as indicated in the text. Growth experiment with Biolector® cultivation system (m2pLabs, Baesweiler, Germany) was performed in 1 mL of CGXII with 222 mM of glucose using FlowerPlate® (m2pLabs, Baesweiler, Germany) at 30 °C, 1100 rpm. Growth experiment with flask was performed in 50 mL of CGXII 222 mM of glucose using 500 mL of baffled flask at 30 °C, 120 rpm. For growth rate calculation, cell growth was monitored online every 10 min for 48 h with Biolector®. Maximum growth rate μ (h−1) was calculated from 20 measuring points of arbitrary unit of the backscattering light (620 nm). Plate image was scanned with Perfection V750-M Pro scanner (Epson, Ludwigshafen am Rhein, Germany). Consumption of glucose was tested with Diabur 5000 glucose test stripes (Roche Diagnostics, Mannheim, Germany) with a detection limit of 0.005% glucose.
4
Zymographic Protein Analysis Protocol
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Single lanes were densitometrically scanned by Epson Perfection V-750 m Pro scanner and data were analyzed using an Image J software [53 ]. Protein extract was mixed with a native rehydration solution (Urea 2 M, Chaps 2%, DTT 0.5%, IPG 1%, trace of Bromophenol blue) and loaded on 11 cm strips pH 3–10. Isoelectric focusing electrophoresis was performed at 15 °C on Multiphor II (Amersham Biosciences, Amersham, UK) with the following protocol: 500 V, 1 mA, 5 W, 1 Vh; 500 V, 1 mA, 5 W, 2500 Vh; 3500 V, 1 mA, 5 W, 10,000 Vh; 3500 V, 1 mA, 5 W, 33,250 Vh. The strips were loaded into 15% T, 2.6% C separating polyacrylamide gels with 0.1% porcine gelatine (1 mm thick). SDS–PAGE was performed at 20 mA/gel for 8 min and 30 mA/gel for about 4 h at 8 °C using Hoefer SE 600 Ruby vertical electrophoresis apparatus. After electrophoresis, gels were treated as previously described for zymography. Electrophoretic gels were scanned by Epson Perfection V-750 m Pro scanner and data were analyzed by PDQuest (Bio-Rad, Hercules, CA, USA).
5
Mandibular Arch Form Classification
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Mandibular models were digitally scanned (Epson® Perfection V750-M Pro Scanner, Seiko Epson Corporation, Nagano, Japan) and a ruler was used for size calibration. The most facial aspect of 13 proximal contact areas around the arch was digitized using AutoCAD software (AutoCAD 2012, Autodesk, Inc., San Rafael, United States). The clinical bracket point for each tooth was located facially via a line perpendicular to that connecting the mesial and distal contact points of each tooth [22 (link), 23 (link), 32 ]. Then, tapered, ovoid, and square arch-form templates (3 M Unitek) were used to classify each case, based on the arch form that provided the best fit to the eight clinical bracket points ranging from the mandibular right first premolar to the left first premolar [33 (link)].
6
Characterization of Nitrogen-doped TiO2 Nanoparticles
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