By: Clarence Halog Emmanuel Egeron Maricar Serrano

What is Spectrophotometer ?
A spectrophotometer measures either the amount of light reflected from a sample object or the amount of light that is absorbed by the sample object. A spectrophotometer consists of two instruments, namely a spectrometer for producing light of any selected color (wavelength), and a photometer for measuring the intensity of light.
http://simple.wikipedia.org/wiki/Spectrophotometer

Kinds of Spectrophotometer: Single beam - A single beam spectrophotometer measures the relative light intensity of the beam before and after a test sample is inserted. Double beam -A double beam spectrophotometer compares the light intensity between two light paths. One path containing a reference sample.

Models of Spectrophotometer:

spectrophotometer model 721

spectrophotometer model 6305 spectrophotometer model 340

Basic Principle of Spectrophotometer Monochromatic light passed through absorbing solution of fixed depth (cuvette) Transmitted light directed to photosensitive 2 Transmitted light directed to photosensitive device Radiant energy converted to electrical energy Instrument adjusted using blank containing all components of unknown solution except the substance being measured Beer Lamberts Law A=bc
http://webmedia.unmc.edu/alliedhealth/CLS/CLS414%2010/SpectrophotometryPrinciplesPartII%2010%20%5BCompat ibility%20Mode%5D.pdf

http://research.ce.udel.edu/~imhoff/cieg337/file_downloads/Spectrophotometry_handouts.pdf

Steps in using Spectrophotometer:

1. WARM-UP: Plug in and turn on (left hand front dial, labeled ZERO in the illustration). Allow about 30 minutes for warm

up.

2. ZERO ADJUST:With no cuvette in the chamber, a shutter cuts off all light from passing though the cuvette chamber. Under this condition therefore, the machine may be adjusted to read infinite absorbance (zero% transmittance) by rotating zero adjust knob (front left on Spectronic 20). Do not touch this knob

again
during the rest of the following procedure.

3. Select the desired wavelength of light at which

4. BLANK ADJUST:Fill the B (blank) cuvette with the solvent used to dissolve specimen (often distilled water). Polish to clean, insert into the cuvette chamber, aligning mark to front. Close chamber cover.

5. Rotate blank adjust knob (front right knob) to adjust absorbance to read zero .
6. Remove blank cuvette, place in plastic test tube rack.

7. READ SPECIMEN: Pour the sample into the S (specimen) cuvette, polish and insert into the chamber, aligning mark to the front.

8. Note that the scale for absorbance is the lower scale on the dial, and should be read from R to L .

http://www.youtube.com/watch?v=20TpmeaTJho

ACTIVITY (Working in Groups). Divide the class into two groups. Half of the class will be working with the spectrophotometer and half the class will be working with the micropipettes.Assign students to work in groups of three. However, during the spectrophotometer activity, students may need to work in larger groups depending on the class size and the number of spectrophotometers available. After 20 minutes, the teacher should have the groups switch.

Sample Journal:
http://www.laserfocusworld.com/articles/2011/02/craic-technologiesspectrophotometer-nondestructively-analyzes-microscopic-samples.html

Craic Technologies spectrophotometer nondestructively analyzes microscopic samples

02/04/2011

The 308 PV UV-visible-NIR spectrophotometer can be added to an open photoport of a microscope or probe station to nondestructively analyze the spectra of many types of microscopic samples. It can acquire spectra by absorbance, reflectance, luminescence, and fluorescence, in addition to highresolution color images, and can be used with NIST-traceable standards. Craic Technologies San Dimas, CA sales@microspectra.com More Products ----PRESS RELEASE CRAIC Technologies Introduces the 308 PV UV-visible-NIR Spectrophotometer for Your Microscope San Dimas, CA (September 1, 2010)-- CRAIC Technologies, the worlds leading innovator of UV-visible-NIR microspectroscopy solutions, is proud to introduce the 308 PV UV-visible-NIR spectrophotometer for your microscope. Designed to be added to an open photoport of a microscope or probe station, the 308 PV microscope spectrophotometer will nondestructively analyze the spectra of many types of microscopic samples. Featuring CRAIC Technologies new Lightblades spectrophotometer technology, the 308 PV can acquire spectra of microscopic sample areas by absorbance, reflectance, luminescence and

microscopes. Applications are numerous and include vitrinite reflectance of coal and spectral analysis of minerals, high resolution colorimetric and relative intensity mapping of flat panel displays and even thin film measurement of semiconductors. Combined with CRAIC Technologies Traceable Standards, which are specifically designed for use with microspectrophotometers and traceable to Standard Reference Materials from NIST, the 308 PV spectrophotometer is a cost effective micro-analysis tool for any laboratory or manufacturing facility. CRAIC Technologies has been an innovator in the field of UV-visible-NIR microanalysis since its founding. We have helped to advance the field of microscale analysis with innovative instrumentation, software, research and teaching. The 308 PV microscope spectrophotometer is the ideal tool for a laboratory or production line due the fact that it can add so many capabilities to a microscope or probe station. It can even be used to upgrade older microspectrophotometers with cutting edge detectors, electronics and software states Dr. Paul Martin, President of CRAIC Technologies. The 308 PV also features our proprietary Lightblades spectrophotometers technology for improved performance and enhanced flexibility. With the addition of advanced spectroscopic, data analysis, film thickness and colorimetric software, this tool is perfect for any environment. The 308 PV spectrophotometer integrates CRAIC Technologies Lightblades spectrophotometer with a sophisticated optical interface hard

Lightblades are spectrophotometers designed and built by CRAIC Technologies specifically for microscale analysis. This new level of sensitivity and stability give us a flexible instrument that is custom designed to add to a microscopes photoport and rapidly acquire high quality spectra from microscopic samples. Sophisticated software, high resolution imaging, permanently calibrated variable apertures and other innovations all yield a new level of sophistication for microanalysis. With high sensitivity, durable design, ease-of-use, imaging and spectroscopic techniques and the support of CRAIC Technologies, the 308 PV is more than just a scientific instrumentit is a solution to your analytical challenges. For more information on the 308 PV microscope spectrophotometer, Lightblades spectrophotometer technologies and the Perfect Vision for Science, visit http://www.microspectra.com/. About CRAIC Technologies: CRAIC Technologies, Inc. is a global technology leader focused on innovations for microscopy and microspectroscopy in the ultraviolet, visible and near-infrared regions. CRAIC Technologies creates cutting-edge solutions, with the very best in customer support, by listening to our customers and implementing solutions that integrate operational excellence and technology expertise. CRAIC Technologies provides answers for customers in forensic sciences, biotechnology, semiconductor, geology, nanotechnology and materials science markets who demand quality, accuracy, precision, speed and the best in customer support. -----