The MEL-BSA (0.25 mg mL−1) antigen was coated in the NC membrane as the test line (T line) and the goat anti-mouse IgG antibodies (0.5 mg mL−1) were coated as the control line (C line) by means of the BioDot XYZ 3000 instrument at densities of 1 μL cm−1. The ICA strip was assembled by pasting the NC membrane on the center of the PVC backing card, and the glass fiber and absorption pads were pasted on both the ends with a nearly 2 mm overlap with the NC membrane. The assembled ICA strip was then dried in a dry oven at 37 °C for 2 h and cut into a width of 3.5 mm by means of an automatic cutter and stored under dry conditions at room temperature until use. The schematic description of MEL-detecting QB-ICA is shown in Fig. 1 .
Xyz3000
Manufactured by BioDot
Sourced in United States
The XYZ3000 is a high-precision laboratory equipment designed for automated liquid handling tasks. It features a multi-channel pipetting system with adjustable volume settings. The device is capable of accurate and reproducible liquid dispensing, aspiration, and mixing operations.
Automatically generated - may contain errors
Lab products found in correlation
8 protocols using xyz3000
1
Rapid Melanoma Detection Assay
2
Immuno-strip Fabrication Protocol
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An immuno-strip was prepared according to the procedure reported in an earlier study20 (link). The strip consisted of four different types of functional membrane pads consecutively connected by partial superimposition. The four membranes were (from the bottom): a polyester pad (4 × 17 mm) for medium application, a glass fiber membrane (4 × 10 mm) as a medium reservoir, an NC membrane (4 × 25 mm) for signal generation, and cellulose chromatography paper (4 × 15 mm) for absorption. The signal generation pad was made by dispensing (1 μL/cm) the capture antibody (clone 560 for single pair, or clones 560 and M18 for double pairs; 1 mg/mL each) and goat anti-mouse IgG (0.1 mg/mL) in PBS onto the respective sites of the NC membrane using a micro-dispenser (a non-contact type; Bio-Dot, XYZ 3000, Irvine, CA, USA). The membrane was dried at 37 °C for 1 h. Finally, the prepared pads were arranged to a width of 4 mm, and each contiguous membrane pad was partially superimposed and fixed onto a plastic film using double-sided tape. A laminating film (3 × 23 mm) was used to cover the surface of the signal generation pad except for the lateral side, which received the horizontal absorption pad (see below)19 (link).
3
Automated Colloidal Gold and Fluorescence Test Strip Analysis
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The XYZ3000 dispensing platform was used for scribing, and the CM2000 guillotine cutter was used for cutting test strips (BioDot, Irvine, CA, USA). A F-4500 fluorescence spectrometer (Hitachi, Tokyo, Japan) was used to detect the test strip data. The colloidal gold test strip results were read using a GY-610 colloidal gold test strip reader (Henan Guanyu Instrument Co., Ltd., Zhengzhou, Henan, China). The FIC-S2011 dry fluorescence immunoassay analyzer (Suzhou Helmen Precise Instruments, Suzhou, Jiangsu, China) was used to read the fluorescent strip data.
4
Colloidal Gold-Based SARS-CoV-2 Antigen Test
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Colloidal gold suspension was prepared by reducing gold chloride with citrate. The colloidal gold was conjugated to recombinant protein rS-RBD. Briefly, 1 mg of rS-RBD was added to 100 mL of the colloidal gold suspension. After a 30-min reaction, conjugation was blocked using 10 mL of 10% bovine serum albumin (BSA) for 15 min. The colloidal gold conjugate collected (centrifugation at 12,000 rpm, 30 min, and 4 °C) was resuspended in PBS containing 0.1% BSA and 0.1% Tween-20.
The colloidal gold and rS-RBD (0.5 mg/mL) conjugate was applied to a conjugate pad (glass fiber) (30 μL/cm, dried at 37 °C, 3 h). Using a dispenser (XYZ3000; BioDot, Irvine, CA), rS1 and the secondary polyclonal antibody (2 mg/mL) were coated onto the nitrocellulose membrane as the test and control lines, respectively, at a dispensing rate of 1.0 μL/cm. The membrane was then dried at 37 °C for 1 h. Finally, the nitrocellulose membrane, conjugate pad, sample pad, and the absorbent pads were assembled and cut into 4-mm strips.
The colloidal gold and rS-RBD (0.5 mg/mL) conjugate was applied to a conjugate pad (glass fiber) (30 μL/cm, dried at 37 °C, 3 h). Using a dispenser (XYZ3000; BioDot, Irvine, CA), rS1 and the secondary polyclonal antibody (2 mg/mL) were coated onto the nitrocellulose membrane as the test and control lines, respectively, at a dispensing rate of 1.0 μL/cm. The membrane was then dried at 37 °C for 1 h. Finally, the nitrocellulose membrane, conjugate pad, sample pad, and the absorbent pads were assembled and cut into 4-mm strips.
5
Lateral Flow Assay for Anthrax Detection
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mAb 8B10 was passively absorbed to 40 nm colloidal gold particles at 1.5 μg/ml, concentrated to OD 6 and sprayed onto a conjugate pad at 6 μl/cm using a BioDot XYZ3000. For the test line, 0.20 mg/ml 8B10 was sprayed at 1 μl/cm onto nitrocellulose membrane. An additional control line reactive with gold conjugate was included to verify proper flow between membranes. Assays were housed in a plastic cassette for ease of use. Cassettes provided a port for sample application and test and control lines were framed by a viewing window.
The limit of detection (LOD) of the LFA constructed with mAb 8B10 was determined using purified B. anthracis Pasteur PGA diluted in pooled normal rabbit plasma and normal human serum. Twenty microliters were applied into the sample port and then followed with 2 drops of a chase buffer. After 15 min, blinded assays were read by four different individuals. Plasma samples from infected rabbits were evaluated for presence of PGA in a similar fashion. Results were reported as positive if at least 3 out of 4 evaluators deemed the assay to be positive.
The limit of detection (LOD) of the LFA constructed with mAb 8B10 was determined using purified B. anthracis Pasteur PGA diluted in pooled normal rabbit plasma and normal human serum. Twenty microliters were applied into the sample port and then followed with 2 drops of a chase buffer. After 15 min, blinded assays were read by four different individuals. Plasma samples from infected rabbits were evaluated for presence of PGA in a similar fashion. Results were reported as positive if at least 3 out of 4 evaluators deemed the assay to be positive.
6
SERS-LFIA Biosensor Fabrication
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The SERS-LFIA biosensor consists of an absorption pad, an NC membrane, a conjugate pad, and a sample pad as demonstrated in Scheme 1 C. Each component was installed on a plastic adhesive backing plate in sequence. The two test lines were separately sprayed onto 0.5 mg/mL of anti-human IgM and 0.6 mg/mL anti-human IgG, whereas the control line was coated onto 0.5 mg/mL of S protein antibody. All three lines were spaced approximately 5 mm apart on the NC membrane via a line dispensing instrument (BioDot XYZ3000). The fabricated NC membrane was dried at 37 °C for 1 h. Finally, the assembled test strips were cut into 3.5 mm width using a programmable cutter (Zeta Corporation, South Korea) for future use.
7
Lateral Flow Strip Fabrication Protocol
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The LFS consists of several key components, such as a sample pad, a filter pad, a conjugate pad, a chromatographic membrane, an absorbent pad, and a baseplate. The QD-CRP135 mAb conjugates and the QD-14C12 mAb conjugates were evenly sprayed onto a pad to make a conjugate pad by XYZ3000 (Bio-Dot). 14A2 mAb was sprayed onto the processed chromatographic membrane to form detection line 1, CRP was fixed to form detection line 2 and goat anti-mouse IgG was fixed to form the quality control line. The detection lines and quality control line are 1.5-mm-wide with 4 mm of space between two different lines. After the pads were laminated, the LFS was cut vertically with a slitter into 5-mm wide, sealed in dry bags and stored at 4 °C.
8
Nitrocellulose Membrane Protein Antigen Assay
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Protein antigens were sprayed on a nitrocellulose membrane (thickness of the membrane: 125–155 μm; pore size: 10μm, capillary flow time: 110–165 [s/4 cm]) at a concentration of 2 mg/ml (in phosphate buffer 10mM, pH 7.3) by a dispenser system (BioDot XYZ-3000 dispensing platform, 1 μL protein solution/cm). Dipstick kits were stored at 4°C.
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