Olympus Fluoview Resource Center_ Interference Filters for Fluorescence Microscopy

8/2/2019 Olympus Fluoview Resource Center_ Interference Filters for Fluorescence Microscopy

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Theory of Confocal Microscopy

Interference Filters for Fluorescence Microscopy

T - , , . F , , , , . T interference filters , .

C , , , . I - - , -- , , . T - , , ,

- . H , . V , .

H . M . I

-- , .

I . T - - , ,


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. T , - . B , /, . A , , .

T , , , cube block. T excitation , dichromaticmirror(beamsplitter), emission (barrier) , F 1. E , . C . I, , , .

A

F 1. T 45- . T (F 1()).L , . T F 1(). D , (380-420 510-560 ). N , 400 . T . A,

, .

I F 2 . T surface flatness (F 2()) , ( 550 ). W

. T , . T transmitted distortion (F 2()), . C , . A wedge (F 2()), - - .I . F , - . I , - .

T


3/17

. T . I - - , , - , . I , 40 (- -; FWHM), . A . A

( ), . P .

D - , . E - , . I , , , , . P --

.

T () 45 ( F 1). T, dichroic mirror, . D , , . P 45- , 90- . S, , .

T . T , , .B , , . I - , . P 1 - , 10 ( ), 1

( ). I ,

, .T 100- , . S, . I , , - .

T , , . T , . T .

T , . T, 5.5 .


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I , , . I , , . L / , , , - . T --

. I , : 1 - 1 . I , , .

P F 3 (F 3()) (F 3()) . T 20- 480- , . T - 500 520 , . U . T 30- (F 3())

(F 3()). H, (40 ) .

Interference Filter Principles

T - . E , - ,- , . S , , , , , , . T- ,

, . C, , . U- , , , , , .

I - . T . T , - .I

. I , , .

R - , , , . W , 4


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. , , 16 ( ). A - . F, . T ( ) .

T . R , , ( ) . W , , ( F 4). T (i) (r)

n(1) n(2), F 4, S' L:

n1 sin(i) = n2 sin (r)

W (i r), :

Rp = [(n2 cos(i) - n1 cos(r)) / (n2 cos(i) + n1 cos(r))]2

Rs = [(n1 cos(i) - n2 cos(r)) / (n1 cos(i) + n2 cos(r))]2

R(p) , p- , R(s) , s- . A , , (R) s p , :

R = Rp = Rs = [(n2 - n1) / (n2 + n1)]2

C , ( 1.0) 1.5, (R) 0.04 - , 0.08 . I , 16 0.96 16 , 52 ( 48- ). T (n(1) 1.0), n(2). T . A , , ,

F 4(), n(2) n(1), 180 .

T , ,


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. , . W , , , .

W , - . T , , Fabry-Perot (F 5()). T

. T - , . T , n(1),n(2), n(3), F 4(). T t, - .

I F 4(), , Fresnel loss ( F reflection, A F).T (R), F (n(2) - n(1)) . A , F . H, ( ), ,

. T , , , , .

I - , n(2), , - - , F 4() ( ). T . B n(2) n(1), ( - ) 180- . I ( , t, ,

n(2)) - (l/4),

- 180- (l/2) . I , 180- . T - - 180- , . W , - -


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., , - .

T - . A , - ( ), 180- . T , , , n(2)

n(3), 180- (l/2)

-- (l/4) . C

( 180- ), - , .

B - , .

A , - - . T , , . C - , -, 180 . C, , . O -. D - - 360 ( ), absentee . F , .

Interference Filter Design

T - . W , . T ( 180 ), , . W , . T- ( - ) . S , .P - , .

C


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, , . M - . B - , . A - , ( ) , - . P F 6 .I ,

(CWL) cut-on cut-off () ( ) , . T ( ) F 6 .

A , - - , , . Tquarter-wave stack reflector( F 5()) , - .T , principal wavelength . T - F 5() - ( 2.35) ( 1.35) , 550 .

A ( ringing) , - . A , . T . Q- , , - .

N () F-P , . I F-P ( ; F 5()), ()

. E . P , , . T , , , (n 1.0) .

T - F-P, cavity single-cavity coating. T, - - ( F 7). I - , - ( ). T -


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- . T , -, . L

. T . P , , , , . A , ,

- -.

A . A , . E - , - (F 7). M-- (MDM) , - . I F-P , - -.

T - , , ( F 7 8). A - period, .T - .T () - . F 7 - . T - .C, , - ( F 9), . T G L ,

.

I - , - - H, - - L (F 7 8). V . O, . Z - , , , - . T - - , () - - .

A , - --


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. . A , , . W , . I , .B - - S'

.

T bleed-through . I , . T - , . F 9() , - , 435 460 , . W , - -

. I - - , . F S', .

I , . T . E , . A , attenuation range - . F, attenuation level . A , .

A , . C, . F, ' , .

I , - F 7, ( ) - . F F 7, - . I , 0.01 . O,

.

E

. A . S ( F 7). T- , , . I


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, .

A , - , . I

, , - . F, , .

I ( ), - .P , . S , , , , , "" .F 10 . N

.

M . O - - .T - . T 10 60. I , - 45 85 .

T - blocking optimized blocking complete. F - . A

. T - , .

M ( ). S : , , , . T

( ), . T . F 25 , . T , ,


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. , , . A- - 45- . T ( F11).

T

( ) . I () , . T , . W ( ), - . C , 665LP () (50 665 ) 605LP 45- .

T . B ,

( 25 ):

l = l0 [1 - (n0/n)2 sin2()]1/2

l() , , l(0) ( ). T

( n(0); 1.0 ) ( n) .

T , . W , - , 1.45 2.0, . T . I - ( ) , - ( ) . I ,

, . I ( 30 ), , .


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- , . A , ; , . A , (p-polarization) (s-polarization). F 11 0- , 45- ,

45- . S - - . T 45- , . A 45- , , -, - .

T , - , , . H, ,

. O , , . I , - . I , , . M .

Interference Coating Production Methods

I . T- , .O

. T - . T , , . I , .

T- , . P (PVD) . P

. V , , PVD .


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R - , . A ( F 12). S . M . I , , . C , , , - ,

- .

A , , . M , . T . D , . L -. A ,

. T . T - - , . T .

A , - . T , , resistive thermal evaporation (boats) , ,

. B , - . T . T , . F, . T , , , . I , - ( ), . T .

T - . Electronbombardment - , , , , - , . T - ( 1 10- ) . I . A, . A - ( reactive

evaporation) .

I , - . R PVD , ion-assisted deposition, ion-beam sputtering, reactive magnetron sputtering.E PVD , ,


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, , , , ( ).

I- - . T , . R ,

. D , , .

I - , - . A , , . A , dual ion-

beam sputtering, ,, - .

A PVD , , . T - , . R . V , .

A -

, . A , , , . T , . S


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, , ), - , I 301 ( ), , BK7 ( ), .

Additional Considerations for Laser Scanning Confocal Systems

T , . A . A,

.A ( ), , , --. T - . T , -

. T , , , , . A , - .

S, - ( 1 -) ( ) . T , . T - ( ) . A - 10 .

I , . B .B , , . D , , , .

T , . T ( - ) . E ,

. T , -- . B -- , .

Conclusion

M . O , . O

, , , .T - - -, -


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.

Contributing Authors

Christopher Hardee, Roy Kinoshita, Travis Wakefield, Robert Johnson - O O, I., 210 M S,

B, V, 05301.

Turan Erdogan - S, I., 3625 B R, R, N Y, 14624.

Kenneth R. Spring - S C, L, M, 20657.

Thomas J. Fellers Michael W. Davidson - N H M F L, 1800 E P D D., T

F S U , T, F, 32310.

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Olympus Fluoview Resource Center_ Interference Filters for Fluorescence Microscopy

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