Fluorescence and Absorption Spectroscopy of Bio-molecules荧光与生物分子吸收光谱
合集下载
- 1、下载文档前请自行甄别文档内容的完整性，平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益，请联系客服反馈,我们会尽快为您处理(人工客服工作时间：9:00-18:30)。
60
G uanine
40
Thym ine
20
0
300 320 340 360 380 400 420 440 460 480 500
W avelength (nm )
FIG. 5
E m is s io n In te n s ity (a .u .)
E m issio n S p e ctra o f D e o xyn u cle o tid e s-m o n o p h o sp h o ra te
Absorption
Absorption is the process in which a substance absorbs or gathers incoming light. Light that is not absorbed is either reflected or transmitted.
F IG .2
FIG. 2
N orm alizedFlourescenceSpectralIntensityofPhenylalanine,Tryptophan andTyrosineinPow der,ExcitationW avelengthsat240nmand265nm
1.0
295
305.5 335
Experimental Results
N o r m a liz e d A b s o r b a n c e
N o rm a liz e d A b s o rp tio n S p e c tra l In te n s ity o f A m in o A c id s o f P h e n y la la n in e , T y ro s in e a n d T ry p to p h a n S o lu tio n in C o n c e n tra tio n 1 0 -3M
electron
E2 > E1
/hbase/hframe.html
Bio-molecules Investigated Table#1
Name
Molecular Formula
Molecular Weight
Molecular Structure
Future Work
• Our future goal is to learn about the Stokes Shift.
• We also would like to conduct experiments that would hopefully lead to better and faster detection of cancerous diseases.
Nor lmiazedItnenisty
0.8
Tryptophan
0.6
Tyrosine
0.4
Phenylalanine
0.2
0.0
260 280 300 320 340 360 380 400 420 440 460 480
W avelengths(nm )
FIG .3
A b s o rb a n c e
High School Summer Research Program
Abstract
We have looked at the fluorescence and absorption of the four bases of deoxyribonucleic acid (DNA), and three amino acids (Phenylalanine, Tryptophan, and Tyrosine). The purpose of this project was to learn about the fundamental science behind disease detection by using fluorescence and absorption. This experiment can be used to find malignant and benign forms of tumors and other cancerous diseases.
a. measuring a certain amount of water (varying from 1mL to 4mL) b. Then the DNA bases were mixed with powder and water
2) The solution was inserted into the vial and then the narrowband filter (optional) was inserted into the fluorescence meter.
3) Tests were run for the fluorescence and the absorption. 4) The results were made into graphs using origin 5.0.
This is a picture of the C-Scan LDS-3 Fluorescence meter we used to measure the fluorescence of the biomolecules.
1 .4 1 .2 1 .0 0 .8 0 .6 0 .4 0 .2 0 .0
220
E le ctro n ic A b so rp tio n S p e ctra o f D p h o ra te (d A M P , d C M P , d G M P , T M P ) in W a te r
N o rm a lize d E m is s io n S p e c tra l In te n s ity o f P h e n yla la n in e , T yro s in e a n d T ryp to p h a n S o lu tio n o f C o n c e n tra tio n 1 0 -3M
360
380
400
420
440
460
W avelength (nm )
F IG . 6
Peak Importance
The peak is important to our research because the peak tells us at what wavelength the bio-molecules fluoresce. Knowing at what wavelength it fluoresces is important because then we have a control sample to compare and contrast whether a tissue is cancerous or not. If the tissue is cancerous, we can determine the intensity or severity of the it.
242.23
C11H12N2O2
204.23
C9H11NO3
181.19
C9H11NO2
165.19
nature/nbt/journal/v19/n4/thumbs/nbt0401_360_F1.gif
Procedures
1) The solution was inserted into a vial by:
E m ission S pectra of D eoxynucleotides-M onophosphorate (dA M P , dC M P , dG M P , TM P ) in W ater
( excitation WL 260 nm) 100
A d e n in e
80
C yto s in e
Deoxyadenosine (A)
Deoxycytidine (C)
Deoxyguanosine (G)
Thymine (T)
Tryptophan
Tyrosine
Phenylalanine
C10H13N5O3 C9H13N3O4 C10H13N5O4
251.24 227.22 595.2
C10H14N2O5
287
305
357
1 .0
0 .8
T rypto pha ne
0 .6
0 .4
T yro s in e
0 .2
P h e n yla la n in e
0 .0
260 280 300 320 340 360 380 400 420 440 460 480
W a ve le n g th (n m )
(d A M P , d C M P , d G M P , T M P ) in P o w d e r
140
312
( excitation WL 240 nm)
120
C ytosine
100
A denine
80
358
G uanine
60
346
40
T hym ine
329.5
20
0
300
320
340
Fluorescence and Absorption Spectroscopy of Bio-molecules
Ivanna Eusebe, RenéCardona and Katherine Gil
Mentor: Ms. Cheng-Hui Liu
Steady State Laser Spectroscopy Lab City College of New York
Fluorescence
When a substance absorbs radiation, atoms in the substance are excited to a higher energy level. The atoms then emit light of lower energy (longer wavelength) and relax to the ground state. The remainder of the energy is released in the form of heat.
D e o xya d o n e sin e T h ym id in e D e o xycytid in e
D e o xyg u a n o sin e
240
260
280
300
320
W a v e le n g th (n m )
F IG . 4
E m is s io n In te n s ity (a .u .)
1 .0
256
273
293
0 .8
T ryp to p h a n
0 .6 T y ro s in e
0 .4 P h e n y la la n in e
0 .2
0 .0
240
260
280
300
320
340
360
W a v e le n g th (n m )
F IG .1
N o rm a liz e d E m is s io n In te n s ity
This is a picture of the Perkin-Elmer Lambda 9 UV/VIS/NIS Spectrometer we used to measure the absorption of the bio-molecules.
The differences between the two machines (not including the fact that one measures fluorescence and the other measures absorption) is that they collect data in different ways. For example if we wanted to see the absorption we would run a background check to make sure that the viles in which we put the bio-molecules would properly allow light to travel through them. Whereas in fluorescence we do not have to go through the previous procedure.
G uanine
40
Thym ine
20
0
300 320 340 360 380 400 420 440 460 480 500
W avelength (nm )
FIG. 5
E m is s io n In te n s ity (a .u .)
E m issio n S p e ctra o f D e o xyn u cle o tid e s-m o n o p h o sp h o ra te
Absorption
Absorption is the process in which a substance absorbs or gathers incoming light. Light that is not absorbed is either reflected or transmitted.
F IG .2
FIG. 2
N orm alizedFlourescenceSpectralIntensityofPhenylalanine,Tryptophan andTyrosineinPow der,ExcitationW avelengthsat240nmand265nm
1.0
295
305.5 335
Experimental Results
N o r m a liz e d A b s o r b a n c e
N o rm a liz e d A b s o rp tio n S p e c tra l In te n s ity o f A m in o A c id s o f P h e n y la la n in e , T y ro s in e a n d T ry p to p h a n S o lu tio n in C o n c e n tra tio n 1 0 -3M
electron
E2 > E1
/hbase/hframe.html
Bio-molecules Investigated Table#1
Name
Molecular Formula
Molecular Weight
Molecular Structure
Future Work
• Our future goal is to learn about the Stokes Shift.
• We also would like to conduct experiments that would hopefully lead to better and faster detection of cancerous diseases.
Nor lmiazedItnenisty
0.8
Tryptophan
0.6
Tyrosine
0.4
Phenylalanine
0.2
0.0
260 280 300 320 340 360 380 400 420 440 460 480
W avelengths(nm )
FIG .3
A b s o rb a n c e
High School Summer Research Program
Abstract
We have looked at the fluorescence and absorption of the four bases of deoxyribonucleic acid (DNA), and three amino acids (Phenylalanine, Tryptophan, and Tyrosine). The purpose of this project was to learn about the fundamental science behind disease detection by using fluorescence and absorption. This experiment can be used to find malignant and benign forms of tumors and other cancerous diseases.
a. measuring a certain amount of water (varying from 1mL to 4mL) b. Then the DNA bases were mixed with powder and water
2) The solution was inserted into the vial and then the narrowband filter (optional) was inserted into the fluorescence meter.
3) Tests were run for the fluorescence and the absorption. 4) The results were made into graphs using origin 5.0.
This is a picture of the C-Scan LDS-3 Fluorescence meter we used to measure the fluorescence of the biomolecules.
1 .4 1 .2 1 .0 0 .8 0 .6 0 .4 0 .2 0 .0
220
E le ctro n ic A b so rp tio n S p e ctra o f D p h o ra te (d A M P , d C M P , d G M P , T M P ) in W a te r
N o rm a lize d E m is s io n S p e c tra l In te n s ity o f P h e n yla la n in e , T yro s in e a n d T ryp to p h a n S o lu tio n o f C o n c e n tra tio n 1 0 -3M
360
380
400
420
440
460
W avelength (nm )
F IG . 6
Peak Importance
The peak is important to our research because the peak tells us at what wavelength the bio-molecules fluoresce. Knowing at what wavelength it fluoresces is important because then we have a control sample to compare and contrast whether a tissue is cancerous or not. If the tissue is cancerous, we can determine the intensity or severity of the it.
242.23
C11H12N2O2
204.23
C9H11NO3
181.19
C9H11NO2
165.19
nature/nbt/journal/v19/n4/thumbs/nbt0401_360_F1.gif
Procedures
1) The solution was inserted into a vial by:
E m ission S pectra of D eoxynucleotides-M onophosphorate (dA M P , dC M P , dG M P , TM P ) in W ater
( excitation WL 260 nm) 100
A d e n in e
80
C yto s in e
Deoxyadenosine (A)
Deoxycytidine (C)
Deoxyguanosine (G)
Thymine (T)
Tryptophan
Tyrosine
Phenylalanine
C10H13N5O3 C9H13N3O4 C10H13N5O4
251.24 227.22 595.2
C10H14N2O5
287
305
357
1 .0
0 .8
T rypto pha ne
0 .6
0 .4
T yro s in e
0 .2
P h e n yla la n in e
0 .0
260 280 300 320 340 360 380 400 420 440 460 480
W a ve le n g th (n m )
(d A M P , d C M P , d G M P , T M P ) in P o w d e r
140
312
( excitation WL 240 nm)
120
C ytosine
100
A denine
80
358
G uanine
60
346
40
T hym ine
329.5
20
0
300
320
340
Fluorescence and Absorption Spectroscopy of Bio-molecules
Ivanna Eusebe, RenéCardona and Katherine Gil
Mentor: Ms. Cheng-Hui Liu
Steady State Laser Spectroscopy Lab City College of New York
Fluorescence
When a substance absorbs radiation, atoms in the substance are excited to a higher energy level. The atoms then emit light of lower energy (longer wavelength) and relax to the ground state. The remainder of the energy is released in the form of heat.
D e o xya d o n e sin e T h ym id in e D e o xycytid in e
D e o xyg u a n o sin e
240
260
280
300
320
W a v e le n g th (n m )
F IG . 4
E m is s io n In te n s ity (a .u .)
1 .0
256
273
293
0 .8
T ryp to p h a n
0 .6 T y ro s in e
0 .4 P h e n y la la n in e
0 .2
0 .0
240
260
280
300
320
340
360
W a v e le n g th (n m )
F IG .1
N o rm a liz e d E m is s io n In te n s ity
This is a picture of the Perkin-Elmer Lambda 9 UV/VIS/NIS Spectrometer we used to measure the absorption of the bio-molecules.
The differences between the two machines (not including the fact that one measures fluorescence and the other measures absorption) is that they collect data in different ways. For example if we wanted to see the absorption we would run a background check to make sure that the viles in which we put the bio-molecules would properly allow light to travel through them. Whereas in fluorescence we do not have to go through the previous procedure.