This application note describes how to use Molecular Devices' chemiluminescent microplate reader to perform the corresponding tests. The combination of instrument and kit is simple and easy to operate, and it exhibits high sensitivity when tested accordingly. Both types of detection kits can be tested in a chemiluminescent microplate reader from Molecular Devices for superior results. Advantage - Sensitive and reliable detection of mycoplasma Introduction Mycoplasma is currently the smallest known prokaryote and is a common source of contamination in mammalian cell culture. Mycoplasma contamination puts a lot of cost to cell culturers because they alter the morphology, vitality, and metabolic characteristics of contaminated cells. Mycoplasma is difficult to find in the medium by a microscope. At present, the pollution for this situation requires a sensitive and reliable detection method. The traditional methods for detection of mycoplasma contamination, such as time-consuming labeling and cumbersome PCR, are difficult to obtain accurate analysis. The MycoAlert Mycoplasma Test Kit and the MycoAertPLUS Test Kit are from Lonza Walkersville, Inc., a reagent based on biochemical reactions coupled with a luminescent microplate reader to detect certain mycoplasma enzyme activities. The presence of these enzymes provides a rapid screening process that allows very sensitive detection of mycoplasma contamination of test samples. As the next-generation test kit, MycoAert PLUS has higher photon output efficiency than the previous generation standard kit, and the chemiluminescent reader has a wider dynamic range, whether it is medium, cell culture supernatant, or water. Can be tested accordingly. The procedure for both kits is identical. The first step is to first add MycoAlert reagent to the cell culture supernatant, and lyse the mycoplasma to trigger the chemiluminescence reaction, as shown in Figure 1. In the second step, the viable mycoplasma is cleaved and the enzyme that reacts with the MycoAlert substrate is released. Catalytic ADP is converted to ATP. By measuring the level of ATP before and after the addition of MycoAlert substrate in the sample, the ratio can be obtained and the presence of mycoplasma is indicated. If these enzymes are not present, the second reading will not increase. (Ratio<0.9 of MycoAlert and Ratio<1.0 of MycoAert PLUS), if the sample is positive, the second detection value is higher than the first time, Ratio>1.2, as shown in Figure 2 for the detection process. material: - MycoAlert Mycoplasma Detection Kit (Lonza cat. #LT07-318) method The basic principles of MycoAlert and MycoAlert PLUS kits are the same, but the concentration range of the two kits is different. The details are as follows. Each kit is configured for reagents on the day of use. The MycoAlert positive control sample dilution relationship is shown in Table 1. 100 μL serial dilutions of MycoAlert positive control samples were added to white 96-well luminescent multi-well plates for 3 replicates. 100 μL of MycoAlert reagent (dissolved in MycoAlert buffer) was added to each well and incubated for 5 minutes at room temperature. The well plates were placed in an illuminated microplate reader and the readings of the selected wells were read using SoftMax® Pro software (reading A). 100 μL of MycoAlert substrate (dissolved in MycoAlert buffer) was immediately added to each well and incubated for 10 minutes at room temperature. The well plate was placed in a luminescent microplate reader for detection and the reading of the selected well (reading B). The instrument settings are shown in Table 2. result We used Lonzede's MycoAlert kit for sensitivity evaluation of all luminescent microplate readers, and Ratio>1.2 for MycoAlert kit quality control samples in the 1:8 dilution ratio, and for MycoAlertPLUS kits The control sample obtained Ratio > 1.2 in the 1:1000 dilution factor relationship. All Molecular Devices' illuminating microplate readers have achieved more than expected sensitivity. We have listed the data values ​​using the SpectraMaxi3x Multi-Purpose Plate Reader and two kits, as shown in Figure 3. Table 3 shows that all Molecular Devices' microplate readers can detect a positive ratio of positive samples to a minimum ratio relationship and can also detect a situation of Ratio > 1.2. in conclusion The results show that all Molecular Devices micro-plate readers with luminosity function can meet the detection requirements of this kit and obtain sensitive detection results; Molecular Devices microplate reader combined with MycoAlert kit can be sensitive and rapid The detection of mycoplasma content ensures timely detection of contaminants and saves valuable time in later experiments. Automatic Massage Foot Bath Machine The automatic massage foot bath machine is a device that uses water and massage rollers to provide a relaxing foot massage. It usually has a basin filled with water and has built-in massage rollers that move and massage the feet. The machine may also have other functions, such as heat therapy, air bubbles, and vibration. Users place their feet in the basin and the machine provides a soothing massage that helps relieve tension and improve foot circulation. Some automatic foot massage machines also come with removable attachments for additional massage options. Automatic Massage Foot Bath Machine,Foot Spa Massageer,Heated Foot Spa,Foot Bath Massage Basin Huaian Mimir Electric Appliance Co., LTD , https://www.mmfootspabath.com
- Simple, easy to operate, quick access to test results
- The test results are simple and clear 
- MycoAlert PLUS Mycoplasma Detection Kit (Lonza cat. #LT07-710)
- MycoAlert Assay Control Set (Lonza cat. #LT07-518)
- 96-well polystyrene solid white plates(Greiner cat. #655075)
- Microplate reader
- SpectraMax i3 multi-function microplate reader
- SpectraMax i3x Multi-Purpose Plate Reader
- SpectraMax Paradigm Multi-Purpose Plate Reader
- SpectraMax M5 multi-function microplate reader
- FilterMax F5 multi-function microplate reader
- SpectraMax L Microplate Reader 
