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All equipment servicing to manufacturer’s specs by Eppendorf factory certified technicians. Calibration, repair, service, spare parts, Compliance Covering - IQ/OQ/PQ, calibration and validations. On-site repairs are available for customers in Sydney, Melbourne and Brisbane. For other areas/States, we offer a workshop service or spare parts.
The new Eppendorf BioSpectrometer is a small, very compact spectrophotometer for measurements in the UV and VIS range. Spectra can be recorded and individual wavelength measurements can be made in a spectral range of 200 nm to 830 nm. Product characteristics:
BioSpectrometer kinetic Only:
BioSpectrometer fluorescence Only:
In addition to standard cuvettes such as the Eppendorf UVette, special microliter measuring cells such as the Eppendorf μCuvette™ G1.0 can be used to measure the most minute volumes. With this option, the Eppendorf BioSpectrometers cover a wide range of concentrations and volumes in addition to standard laboratory methods. Too easy! How the BioSpectrometer works:
 Eppendorf BioSpectrometer® basicThe Eppendorf BioSpectrometer basic offers a wide variety of procedures for the most diverse molecular biology, cell biology and biochemical methods.Eppendorf BioSpectrometer® kineticThe Eppendorf BioSpectrometer kinetic offers all versions of the Eppendorf BioSpectrometer basic as well as a temperature-controlled cuvette shaft (+20°C to +42, 0.1°C increments). Thermoregulation is controlled by an integrated Peltier element. which guarantees extremely accurate temperature control, even over long periods, that allows you to determine enzyme and substrate kinetics directly in the device. No separate accessories are required.
Eppendorf BioSpectrometer® fluorescenceThe BioSpectrometer fluorescence has all of the features of the BioSpectrometer basic, plus the option to determine very low concentrations of biomolecules using fluorescent dyes. In the absorption range, spectra measurements can be made between 200 nm to 830 nm. The fluorescence unit of the BioSpectrometer fluorescence can be used to increase the measuring range by a factor of 1,000, for detecting DNA for example. This means that DNA can be reliably quantified well beyond the common lower photometric detection limit of approx. 2.5 ng/μL, and up to a concentration of 2.5 pg/μL.The fluorescence intensity is determined using an excitation wavelength of 470 nm and emission wavelengths of 520 nm and 560 nm. Well-known dyes for nucleic acid and protein quantification, such as PicoGreen, OliGreen, RiboGreen and NanoOrange, can be measured.
BioSpectrometer fluorescence specifications
UV/Vis BioSpectrometer reference filter set
The UV/Vis BioSpectrometer reference filter set is used to determine the photometric accuracy and wavelength systematic error in accordance with NIST. It is for use with the BioSpectrometer basic and BioSpectrometer kinetic. Product characteristics:
the Eppendorf Uvette® is the perfect cuvette for the BioSpectrometerThe UVette fulfils the highest specifications of modern spectrophotometric analysis in the UV range and can be optimally used for sensitive samples and small volumes. Universal!The new UVette is ideal for use with the BioSpectrometer and the Eppendorf BioPhotometer D30. In other photometers or spectrometers the Uvette is inserted with the aid of an adapter (information on the adapter tab). This adapter aligns the UVette to the corresponding height of the optical light path and also serves as an aperture for optimal light guidance and use of the small measuring volume.Precise!Eppendorf's vast experience in the manufacturing of optical precision parts is evident in every detail. The UVette is subject to our rigorous quality control and test procedures, which safeguard the high-quality standards for which Eppendorf is renowned throughout the world.
Some of the Frequently asked questions about Eppendorf BioSpectrometer® courtesy Eppendorf sp Can I use the BioPhotometer Data Transfer Software also with the BioPhotometer 6131? Which cuvettes can be recommended for the Eppendorf BioSpectrometer? Can the Hellma® TrayCell be used for micro volume measurements in the Eppendorf BioSpectrometer? During the start of the BDTS program I get an error message! What can be the reason? Which cuvettes are recommended for a temperature control in the Eppendorf BioSpectrometer kinetic? How can I create a new "archive"? Is it possible with the new BDTS to create specific user profile? In how many languages will the new BDTS be offered? Can different absorbance spectra be compared directly together in the Eppendorf BioSpectrometer? Will the new BDTS software also work Apple Macintosh computer? What are the system requirements for the BDTS? At what height does the light beam pass through the cuvette in the BioPhotometer? Is the BioPhotometer PC online program compatible with Windows XP? Is the PC online program of your BioPhotometer software package compatible with Windows NT 4.0? Can methods re-imported to the Eppendorf BioSpectrometer? Can I measure the concentration of ring-shaped DNA on the BioPhotometer? Can I use my BioPhotometer filter set for the verification of the BioSpectrometers? Can I measure double-strand RNA on the BioPhotometer? Can I record standard curves on the BioPhotometer? Can I also connect printers other than the Thermal Printer DPU 414 to the BioPhotometer? Can I use the BioPhotometer for protein determination in plasma following the Biuret method? Can I print out data in Excel format over the RS-232 interface of the BioPhotometers? Can the measurement values from the BioPhotometer be transferred to a PC? Does the OD 600 method take place linearly on the BioPhotometer? How often should the BioPhotometer be checked with the Secondary UV-VIS-Filter set? What are the dimensions and weight of the Thermal printer DPU 414? What molecules have an absorption maximum of 230 nm? Which detector is found in the BioPhotometer? What is the smallest filling volume of a cuvette that can be measured on a BioPhotometer? What is the smallest quantity of DNA that can reasonably be measured with the BioPhotometer? What is the smallest quantity of RNA that can reasonably be measured with the BioPhotometer? Which cable is required for connecting the Thermal Printer DPU 414 to the BioPhotometer? How can I clean the cuvette shaft of my BioPhotometer? How can I convert an OD 600 value to cell number/ml? Where can I get my Secondary UV-VIS-Filter set recalibrated?
This is caused by the measurement outside of the upper measuring range (> 3.0 A).
Please check 1. Whether your cuvette is pressed firmly onto the base of the cuvette shaft. The error message "++++" indicates that the cuvette base is blocking the light beam. 2. Whether your cuvette has an optical window at 8.5 mm. 3. Whether you have sufficient blank medium in the cuvette.
Yes, in the method group “Routine” are all methods that also can be found in the BioPhotometer plus. These methods are the detection of nucleic acids, proteins, fluorescent dye labeled biomolecules plus OD600 measurements.
Yes, the software is valid for the BioPhotometer plus as well as for the BioPhotometer 6131!
All cuvettes with standard dimensions can be used (12.5 * 12.5 mm). It is important that at height of 8.5 mm the optical window is transparent for the light beam)? Please check therefore also the specification in the manual (chapter 5.2).
Yes, this is possible. You need an RS232/USB connection cable. This cable is worldwide available by the company VScom.
Negative A230 values are usually caused by interference components in inadequately concentrated DNA solutions. The negative value should be rectified when you use a lesser sample dilution for the next measurement. Please note that you should measure at 260 nm for a minimum absorbance of 0.1 in order to obtain exact results. This value is independent of the measuring device and is based upon the disturbing influence of impurities and particles on the measurement result, which is especially high at A < 0.1.
"Yes, this is possible. To consider the light path for the calculation of the sample concentration, this value has to be entered in the area “check parameters”. Please note, that for the use of the TrayCell a sufficient sample concentration has to be applied: For the available sample concentration we recommend the absorbance ranges (e.g. dsDNA):
0.1 mm = 500 – 5000 ng/µL
0:2 mm = 250 – 2500 ng/µL
1 mm = 50 – 500 ng/µL
2 mm = 10 – 100 ng/µL
These concentration ranges for dsDNA correspond to an absorbance of 0.1 to 1 A.
"
Please note that you should measure at 260 nm for a minimum absorbance of 0.1 in order to obtain exact results. This value is independent of the measuring device and is based upon the disturbing influence of impurities and particles on the measurement result, which is especially high at A < 0.1.
An A260/A230 < 2.0 can indicate impurity of the DNA sample. This is because peptides, aromatic groupes, phenols and carbohydrates absorb at 230 nm.
With contaminated DNA samples you can try to clean the sample through centrifugation (at least. 5 min). Otherwise the sample must be purified again according to the protocol specifications.
Please check, if all systems settings as stated in the manual in chapter 2.3 are correct.
Yes, this is easily possible. However, for absorbance measurement below 220 nm you have to consider the self absorbance of the UVette. For this reason we recommend to carry out measurements in the UVette only above 220 nm.
Yes, this is possible. Click in the "work" area on "ID". Here you can connect you results to a specific series of measurement!
The Bradford reagent is an aggressive substance that attacks the plastic of plastic cuvettes when the exposure time is too long. This reagent should therefore not remain in a plastic cuvette for more than 5 minutes. Fluctuating values could be caused by the fact that the Bradford reagent has already been too long in the cuvette.
Tip: Use glass cuvettes for the Bradford method.
Basically every cuvette with standard dimensions can be used for the BioSpectrometer kinetic (s. chapter 5.2 in the manual). For optimal temperature control it is important that the cuvette has direct contact with the integrated peltier-element. Most adequate cuvettes are made of glass or quartz glass. In chapter 5.3.4 of the manual an overview is shown regarding pre-incubation times for different cuvette shapes.
Yes, this is possible. Therefore you can use in the area "archive" the filter function.
Please check whether the factor 1.000 is found under "Parameter" on the BioPhotometer. It is possible that the factor was accidentally reset to nil when working with the BioPhotometer. In case you continue to have difficulties with measurement, please contact our Application-Hotline: Tel. +49 180 3 66 67 89; email: support@eppendorf.com.
Because of the shape of UVette the temperature transfer is here limited we recommend conventional cells made of glass or quartz glass!
Please check whether:
- the correct concentration unit was selected (e.g. mg/ml was chosen, but the sample has a concentration in the µg/ml range) - an entry with decimal places was made for the standard concentrations. With an entry without decimal places, lesser sample concentrations (e.g. 0.024) cannot be recognized.
No, this data will be exported automatically.
Click top menu bar on "new archive file"!
The life span of the xenon lamp is influenced by neither repeated switching on and off nor by leaving it on in standby over a period of several hours. You can therefore decide for yourself.
The xenon lamp is also only active during the measurement. It then switches over into standby mode.
In the method, "Advanced kinetics”, the measurement of standards can be programmed. This can be used for the measurement of unknown substrate concentrations via enzyme kinetics. Moreover it is possible if a drift of the reagent solution is expected, a “reagent blank” can be measurement. The result of this measurement is than subtracted from the sample measurement. These measurements are in the method "Simple Kinetics" not possible.
Yes, this is possible! Just click on top menu bar on "file" and "settings". In the right window you can apply a new user profile with a new password. Please note that only a new profile can apply if you are login as an administrator.
Please switch the BioPhotometer off and then on again following the installation of the program. The header line will then appear.
The BDTS will only be offered in English language.
Precision measurement should be carried out using a semimicro-quartz glass cuvette (> 350 µl). Clear and clean solutions are suitable as the measuring medium, for example potassium dichromate (zero adjustment to water) or water (zero adjustment to air). The coefficient of variation (CV) should be smaller/equal to 1% at 1 A in line with the technical data for the BioPhotometer. Please bear in mind that the Secondary UV-VIS filter should be used for exact precision measurement and determination of both the photometric and wavelength accuracy.
No, this is due to patent reasons not possible. But you can export the graph data as an Excel file and then reprocess here accordingly.
No, the BDTS will only be available for Windows.
Yes. For each colorimetric calibration saved on the BioPhotometer, all calculated parameters, such as, for example, the ascent of the calibration lines, are automatically filed in the affiliated calibration report. To call up the calibration report press the Function key, select "Calibration report" with the cursor, press the Enter key, select the desired calibration with the cursor, press the Enter key.
No, also this function is due to patent reasons not possible.
Windows XP or Windows Vista.
At 8.5 mm.
The files will be exported as an Excel file.
Yes.
No, this is unfortunately not possible. But it is possible to export the data to an USB-Stick. The data can then be printed from the PC over a local or network printer.
Yes.
A re-import of methods is unfortunately not possible.
Yes. Double-stranded ring-shaped DNA can be measured on the BioPhotometer using the "dsDNA" method, and single-stranded ring-shaped DNA with the "ssDNA" method.
For the BioSpectrometer an additional filter set will be available.
Yes, if you are aware of a factor for double-strand RNA. You should enter this under the RNA method before the measurement under "Parameters". If you don't have a factor, you can carry out a calibration with a familiar dsRNA quantity and then use this to measure the unknown sample.
Yes. This is possible with the Bradford/Bradford micro, Lowry/Lowry micro and BCA/BCA micro methods.
From software version 1.20 on you can also connect other serial and parallel printers (a converter cable is required) besides the Thermal Printer DPU 414 to the BioPhotometer. If you use the BioPhotometer with software version 1.0, you can only connect the Thermal Printer DPU 414.
Because the Biuret method can also be carried out at 562 nm (the absorption behavior of the Biuret reagent is almost identical in the range from 530 to 570 nm), you can measure your Biuret method on the BioPhotometer using the BCA method. Please note that you need only work with a standard for the Biuret method. The measurement of a standard curve is unnecessary.
You can save one standard and one micro method for each protein determination method. This means that you can store two calibration curves for each protein determination method.
From software version 1.20 on you can transfer measuring values from the BioPhotometer to a PC (respectively, in an Excel table) with the help of the BioPhotometer software package and then process them further or print them out.
As of software version 1.20 you can easily and comfortably transfer measurement data into a table calculation program such as Excel using our BioPhotometer software package. The PC online program also enables the use of the familiar printout design from the thermoprinter with a PC printer.
Please check whether
1.The absorbance of your DNA sample is greater than/equal to 0.1 (please note that regardless of the device used, absorption measurements of nucleic acids only deliver reliably reproducible results at absorbances of greater than 0.1. This is based upon the disturbing influence of impurities and particles on the measurement result, which is especially high at A < 0.1.) 2. Your sample was sufficiently mixed (only careful mixing of the sample can guarantee consistent measurement values) 3. There is an impurity in your sample. An indicator of a possible impurity is the relative ratio of A260/A280 or A260/A230, which with pure DNA should amount to app. 2.0 or 2.5 at pH 7-8.5. Carbohydrates, aromatic groups, phenols and peptides have an absorption maximum of 230 nm, proteins of 280 nm. Additional literature: "Tipps and Tricks for photometric quantification of nucleic acids".
You can of course vortex and centrifuge your sample and then transfer it to a cuvette. Please make sure that the sample is transferred to the cuvette air bubble-free .
You fill a cuvette with water and measure it as blank. Now fill another cuvette with water as well and measure this cuvette as a sample. The difference in absorption between the two cuvettes can now be identified as positive or negative absorption at the individual wavelengths. The most reliable method is to repeat the sample measurement up to three times and then to calculate the cuvette error as the mean value of the absorption differences measured.
For purposes of data documentation, either a printer or a PC can be connected to each BioPhotometer. It is also possible to have the photometric systematic (trueness) and photometric random error (precision) of your device inspected by one of our service partners. For purposes of testing directly, we offer the user a Secondary UV-VIS-Filter.
This measuring method is principally non-linear, when one observes the absorption curve over wide OD ranges and dilutions. The values are dependent on both the organism (size, shape) and the device (light path geometry, ratio of measured stray light radiation). A correct procedure includes the generation of a calibration graph for each organism and device by determining the actual number of cells under a microscope. When the curve shape is known it is then possible to measure in ranges that are almost linear.
We recommend checking the photometric systematic error (photometric trueness) and the wavelength systematic error (wavelength trueness) once a year using the Secondary UV-VIS-Filter set.
Factor F is calculated from the molar absorbance coefficient of a molecule in solution and the optical path length of the cuvette used (see Beer-Lambert law). The absorption behavior of nucleic acids is influenced by the aromatic rings of the bases. Here the molar absorbance coefficient of a double-stranded nucleic acid molecule with which the bases are in close contact with each other, is smaller than of a single-stranded nucleic acid molecule. It thus follows that single-stranded nucleic acid molecules have a higher rate of absorption than double-stranded molecules (Sambrook et al. 2001. Molecular Cloning 3rd edition, A8.20).
With an optical path length of 10 mm and under neutral to slightly alkaline measuring conditions an optical density of 1 at 260 nm thus corresponds to approx. 50 µg/ml of double-stranded DNA, 37 µg/ml of single-stranded DNA, 40 µg/ml of RNA and appr. 30 µg/ml of oligonucleotides.
Dimensions (WxHxD) = 160 mm x 70 mm x 170 mm; Weight app. 400 g.
Carbohydrates, aromatic groups, phenols and peptides have an absorption maximum of 230 nm.
Wilfinger et. al* generally recommend using 1 - 3 mM Na2HPO4, pH 8.0 - 8.5 as the measuring medium for nucleic acids. For DNA measurements, our experience shows that 10 mM Tris-HCl, pH 8.0 or a TE buffer, pH 8.0 are also suitable.
*Wilfinger W.W., Mackey K. and Chomczynski P. 1997. Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. BioTechniques 22: 474-481.
Silicium photo diodes are used as detectors in the BioPhotometer.
That depends upon the cuvette chosen. We recommend using the UVette with a minimum volume of 50 µl.
The BioPhotometer should not be used to measure an absorbance of less than 0.05 at 260 nm. This OD value corresponds to a dsDNA concentration of app. 2.5 µg/ml (= app. 125 ng dsDNA in a 50 µl preparation).
Please note that you should measure at 260 nm for a minimum absorbance of 0.1 in order to obtain exact results. This value is independent of the measuring device and is based upon the disturbing influence of impurities and particles on the measurement result, which is especially high at A < 0.1.
The BioPhotometer should not be used to measure an absorbance of less than 0.05 at 260 nm. This OD value corresponds to a RNA concentration of app. 2 µg/ml (= app. 100 ng RNA in 50 µl preparation).
Please note that you should measure at 260 nm for a minimum absorbance of 0.1 in order to obtain exact results. This value is independent of the measuring device and is based upon the disturbing influence of impurities and particles on the measurement result, which is especially high at A < 0.1.
The Thermal Printer is delivered equipped with an appropriate cable. Replacement cables are available from us upon request.
In general, the following applies: F = 1: (e x l) where F = factor [(µg/ml)-1 cm-1], e = molar extinction coefficient [M-1 cm-1], l = optical layer thickness of cuvette [cm]. For small molecules like oligonucleotides, for example, the correct extinction coefficient is determined from the base composition. As the concentration of oligonucleotides is commonly reported as mmol/liter, a millimolar extinction coefficient (E) is conventionally used in the Beer-Lambert equation by means of: E = A (15.3) + G (11.9) + C (7.9) + T (9.3)
A, G, C and T here stand for the number of corresponding bases in this oligonucleotide, the numbers in brackets for the molar extinction coefficient of any deoxynucleotide at pH 7.0.
Source: Sambrook et. al. Molecular Cloning (2001).Third Edition, A8.21
Dust in the cuvette shaft can be carefully removed with a cotton tip. If the cuvette shaft is heavily soiled, for example with dried-on buffer residues, the cotton tip should be carefully soaked in 40% methanol beforehand.
Conversion is possible through calibration graphs or counting. With measurements of Escherichia coli, for example, it is possible to estimate roughly that an OD 600 between 0.5 and 1.0 corresponds approximately to a cell number between 1 x 108 and 1 x 109.
It is 3 m long.
It is 2 m long.
Please send your Secondary UV-VIS-Filter set to POCD Scientific (POCDS).
The 260 nm and 280 nm filter samples are for the testing of the wavelength accuracy, the A1, A2 and A3 filter samples for testing of the photometric accuracy (systematic and random error) of all wavelengths. The values measured with these filters must lie within the given limits. The limiting values are contained in a table found on the inside of the lid of the filter box.
Some video showing you the Eppendorf BioSpectrometer® in action click link to make video visible, click again to vanish video
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www.pocdscientific.com.au/Eppendorf_BioSpectrometer_temperature-controlled_spectrophotometer.php updated June 18th,2013 ©2013 POCD Pty Ltd, T/A POCD Scientific ABN 93 067 939 824 product information, images and video gratefully used with permission of manufacturers. |