ECEN 4616/5616 4/8/2013 Design of a 10x, 0.25 N.A., Microscope Objective We intend to construct the objective from two separated achromatic doublets. Researching the literature shows that such objectives are essentially limited to N.A.s of 0.25 or less. Higher N.A. can be achieved, but with significant complication to the optical design. The following table from a microscope informational website (http://micro.magnet.fsu.edu/primer/anatomy/numaperture.html ) confirms that this design form is limited to ≤ 0.25 N.A.: Objective Numerical Apertures Magnification Plan Achromat (NA) Plan Fluorite (NA) Plan Apochromat (NA) 0.5x 0.025 n/a n/a 1x 0.04 n/a n/a 2x 0.06 n/a 0.10 4x 0.10 0.13 0.20 10x 0.25 0.30 0.45 20x 0.40 0.50 0.75 40x 0.65 0.75 0.95 40x (oil) n/a 1.30 1.00 60x 0.75 0.85 0.95 60x (oil) n/a n/a 1.40 100x (oil) 1.25 1.30 1.40 150x n/a n/a 0.90 “Standard” Microscope Layout: Eyepiece “Artistic” Ray Trace Objective pg. 1 ECEN 4616/5616 4/8/2013 The “standards” for microscope design have historically varied widely. (For a brief description of this history, you can find websites like this one: http://www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopyuk.org.uk/mag/artapr10/rp-compat.html) During the last half of the 20th century, however, a (sort-of) standard mechanical layout was as follows: Tube length: 160 mm Tube diameter (inner): 23 mm. (Since the eyepieces slip into the tube, the maximum clear aperture of the eyepiece optics is considerably less than this.) Bottom of tube to focal plane: 36mm (Japanese) or 45mm (German). (This is only of importance when you have multiple objectives mounted on a turret below the tube – they should all be the same standard!) Specifications: 10X: The objective is to produce a real image of the specimen that is magnified 10 times at a distance of 160 mm (standard microscope tube length) behind the mounting shoulder of the objective: 160 mm Objective 10X Image Eyepiece Specimen pg. 2 ECEN 4616/5616 4/8/2013 0.25 NA: The sine of the half-angle of the light cone from the specimen to the objective is to be 0.25 (14.48 degree half angle cone). aA ==30 dego 14.48 a Since the resolution on the specimen is of prime importance, we will design the objective backwards from the way it is to be used – rays will be traced from the image to the specimen. This way, the PSF and MTF analysis by the ray trace program will directly indicate the resolution achieved at the specimen. We can think of this as “imaging the detector onto the specimen”. Initial Paraxial Layout: h u l u’ l’ K We are given that l 160mm , 𝑢′ = 0.26, and M 1 . 10 Using the paraxial equations, we can calculate that: 𝑀= 1 𝑙′ 𝑙′ 1 𝑙 ⇒ 𝑙 ′ = 16𝑚𝑚 1 = 𝑙 + 𝑓 ⇒ 𝑓 = 14.545𝑚𝑚 ℎ 𝑢′ = − 𝑙′ ⇒ ℎ = 4.16𝑚𝑚 𝑢 𝑀 = 𝑢′ ⇒ 𝑢 = 0.026 (tan 1.49𝑜 ) and 𝑓 𝑓 𝐹# ≡ 𝐷𝑖𝑎𝑚𝑒𝑡𝑒𝑟 = 2ℎ = 1.7 pg. 3 ECEN 4616/5616 4/8/2013 Note that the F/# is faster than the practical limit for achromats (~ F/2 -- determined by scanning the various lens catalogs in Zemax). This confirms our decision to divide the design into two achromats. Note on resolution: The maximum resolution of the objective (period of cutoff spatial frequency) will be 𝑝 = 𝜆 ≅ 1µ𝑚 for green light (0.5µm wavelength). Hence, the spotsize in the 10X 2𝑁.𝐴. magnified image will be 10µm. This is matched well for direct detection using a digital detector with no more than 10µm sized pixels. The maximum resolution of the “standard” human eye is 1 arc min (0.0003 rad). At the standard near-vision distance of 250 mm, this corresponds to a spot size of 0.003*250 = .075mm, or 75 microns. A 10x eyepiece will then reduce the just-resolvable spot size to 7.5 microns, slightly less than the smallest resolvable spot in the objective’s image. A slight excess of “empty resolution” is desirable, as it prevents the user from straining at the limit of their eye’s resolution. Thus, a 0.25N.A., 10X microscope objective is well matched directly to a reasonable digital detector, and to the Human eye using a 10X eyepiece. Revised Paraxial Layout: u2 h1 u1 l h2 d1 K1 K2 u3 d2 For now, we arbitrarily set d2 = 10mm. This may not be optimum, but the optimizer can change it later, while maintaining an adequate working distance. We also divide the ray’s angle change equally between the two lenses. We now calculate the additional variables. Note that u1, l, h1, and u3 are still the same. u1 tan(1.490)=0.026 u2 u2 = tan(3-1)/2 = tan(-80)= -0.14 u3 -0.26 = tan(-14.60) h1 h1 = -u1l = 4.16 mm h2 h2 = -u3d2 = 2.6 mm l -160 mm d d1 = (h2-h1)/u2 = 11.14mm K1 K1=(u1-u2)/h1 = (25.06 mm)-1 K2 K2=(u2-u3)/h2 = (21.67 mm)-1 pg. 4 ECEN 4616/5616 4/8/2013 Zemax Reality Check: (2x vertical stretch) Layout 4/8/2013 Total Axial Length: 181.14000 mm Vertical scale stretched by 2.0000 X 10XMicro_paraxial.zmx Configuration 1 of 1 Calculate Achromat Starting Designs: We now need to replace the paraxial lenses with achromats. Pick some likely glass combination that has a fairly large difference in V-numbers. It is good to stay away from “exotic” glasses at first, as they can greatly increase the fabrication cost. A good guide for achromat glasses is Edmund Optics inexpensive achromats, accessable from the Lens Catalog dialog box (click on the “Len” tab on the menu bar). pg. 5 ECEN 4616/5616 4/8/2013 An easy way to see what these lenses are made of is to insert a bunch of dummy surfaces at the top of the Lens Data Editor, then select a lens and hit “Insert”: pg. 6 ECEN 4616/5616 4/8/2013 Highlighting a glass in the LDE, then clicking on the Glass Catalog button (“Gla”) will bring up the data on that glass: Since our lenses are still fairly fast, we would like two glasses with fairly high indices and a large difference in V-numbers. After looking at a number of achromats and glasses, we pick: Glass # 1) LAK21 2) SFL6 Index 1.640495 1.805182 Vd 60.1019 25.3939 Recall that the paraxial formulas for an achromatic doublet are: K1 K2 K and VK VK K1 1 and K 2 2 V1 V2 V1 V2 Putting the glass and paraxial power values into these formulas (glass 1 = LAK21, glass 2 = SFL6), we get: K1=1.731644 K, and K2 = -0.731644 K pg. 7 ECEN 4616/5616 4/8/2013 Using the thin lens formula, K (n 1)(c1 c2 ) , and assuming, for a starting point, that the positive lens of the doublet is equiconvex (c2= -c1), we get the starting point lens parameters: Paraxial lens 1 2 f f1 R1=-R2 f2 R2 25.9 mm 21.87 mm 14.956885 mm 12.6296 mm 24.086 mm 16.178mm -35.34 mm -29.89 mm -156.8665 mm -104.4089 (Where we have used K=1/f, c = 1/R to make the values easier to enter into Zemax.) Zemax Optimization: Entering the first achromat into Zemax, and re-focusing the system: pg. 8 ECEN 4616/5616 4/8/2013 The lens is acting as an achromat: But, the performance is not great: TS Diff. Limit TS 0.0000 mm TS 6.0000 mm 1.0 0.9 Modulus of the OTF 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 100 200 300 400 500 600 700 800 900 1000 Spatial Frequency in cycles per mm Polychromatic Diffraction MTF 4/8/2013 Data for 0.4861 to 0.6563 µm. Surface: Image 10XMicro_L1_P2.ZMX Configuration 1 of 1 pg. 9 ECEN 4616/5616 4/8/2013 There is a large amount of Spherical Aberration Present: STO Spherical 2 Coma 3 Astigmatism 4 Field Curvature 5 Distortion Axial Color SUM Lateral Color Seidel Diagram 4/8/2013 Wavelength: 0.5876 µm. Maximum aberration scale is 0.05000 Millimeters. Grid lines are spaced 0.00500 Millimeters. 10XMicro_L1_P2.ZMX Configuration 1 of 1 A merit function was constructed to hold the 3rd order spherical and chromatic focal shift to low values – this mimics the old technique of designing by making small changes while keeping 3rd order aberrations low. It helps keep simple systems from assuming weird proportions (as long as the degrees of freedom are available to limit the aberrations): Both wavefront and spot size default functions were tested – the wavefront worked better. pg. 10 ECEN 4616/5616 4/8/2013 The optimization results were good: TS Diff. Limit TS 0.0000 mm TS 6.0000 mm 1.0 0.9 Modulus of the OTF 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 100 200 300 400 500 600 700 800 1000 900 Spatial Frequency in cycles per mm Polychromatic Diffraction MTF 4/8/2013 Data for 0.4861 to 0.6563 µm. Surface: Image 10XMicro_L1_P2_Opt.ZMX Configuration 1 of 1 STO Spherical 2 Coma 3 Astigmatism 4 Field Curvature 5 Distortion Axial Color SUM Lateral Color Seidel Diagram 4/8/2013 Wavelength: 0.5876 µm. Maximum aberration scale is 0.02000 Millimeters. Grid lines are spaced 0.00200 Millimeters. 10XMicro_L1_P2_Opt.ZMX Configuration 1 of 1 (Note the scale on the Seidels is 40% of the unoptimized scale.) pg. 11 ECEN 4616/5616 4/8/2013 Layout 4/8/2013 Total Axial Length: 35.31380 mm 10XMicro_L1_P2_Opt.ZMX Configuration 1 of 1 Continuing on and replacing the second paraxial lens: Layout 4/8/2013 Total Axial Length: 38.53689 mm 10XMicro_L1opt_L2_nopt.ZMX Configuration 1 of 1 Re-optimizingThe lens was re-optimized, after adjusting the merit function to include the second lens. At first, just the second lens was optimized, but this didn’t produce satisfactory results, so both were optimized together: pg. 12 ECEN 4616/5616 4/8/2013 Results, however, were still disappointing: TS Diff. Limit TS 0.0000 mm TS 6.0000 mm 1.0 0.9 Modulus of the OTF 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 100 200 300 400 500 600 700 800 900 1000 Spatial Frequency in cycles per mm Polychromatic Diffraction MTF 4/8/2013 Data for 0.4861 to 0.6563 µm. Surface: Image 10XMicro_L1opt_L2_opt.ZMX Configuration 1 of 1 The layout diagram showed that the first lens had changed a lot, but the second had not. Possibly, it was a mistake to optimize the first lens in the presence of the second paraxial lens – both lenses, perhaps, should have been optimized together: Layout 4/8/2013 Total Axial Length: 42.54483 mm 10XMicro_L1opt_L2_opt.ZMX Configuration 1 of 1 pg. 13 ECEN 4616/5616 4/8/2013 The obvious next thing to try would have to been to back up, put both starting lenses into the design, then optimize that. However, it was late and I wanted to go to bed, so I decided to turn the “Hammer” optimization algorithm loose on the system to see if it could recover by itself. After about 30 minutes, with only moderate results (to be sure, I should have left it going all night), I decided to allow the Hammer algorithm to substitute other Schott glasses, in case the ones I had selected were somehow limiting the results (instead of my optimizing order!). This is done by right-clicking on the glass name in the Lens Design Editor and filling in the following dialog box: Often, it would be wise to limit the glass selection to less than a full catalog (which contains obsolete glasses). This can be done easily by following the directions in the manual on creating new glass catalogs. Make a copy of the Schott catalog, for example, re-name it – say “Schott_Preferred” – and delete all the obsolete glasses. You can also create a catalog to match the glass inventory of a manufacturer. Overnight run of “Hammer” with glass substitution resulted in a good system: pg. 14 ECEN 4616/5616 4/8/2013 pg. 15 ECEN 4616/5616 4/8/2013 The glasses had all been changed – it was evident that the system was not achromatic per lens (see the Seidel Diagram), but was dividing the aberration corrections between the lenses. While this resulted in a good system (better than the one in Zebase!), the division of aberration correction will interfere with any attempts to convert this lens to use of stock achromats. It would be better to do the optimization with additional operands in the MFE that constrained each lens to be individually corrected. pg. 16 ECEN 4616/5616 4/8/2013 Previous attempt at Off-The-Shelf (OTS) microscope lens: Doing this is tedious, as there are no “Substitute” solves that can try different catalog lenses in Zemax – it all has to be done manually. Starting from a similar design, catalog lenses were substituted for the optimized lenses, and the system re-optimized (except for the catalog lenses, of course!). This process only produced reasonable results after the N.A. was reduced to 0.18, and a third lens was added: Layout Stock lens 10x microscope objective, 0.125 NA 4/8/2013 30.39202 mm Total Axial Length: MDE_04_OTS.zmx Configuration 1 of 1 Within those constraints, however, the results were fairly good: pg. 17 ECEN 4616/5616 4/8/2013 TS Diff. Limit TS 0.0000 mm TS 5.0000 mm 1.0 0.9 Modulus of the OTF 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 74 148 222 296 370 444 518 592 666 740 Spatial Frequency in cycles per mm Polychromatic Diffraction MTF Stock lens 10x microscope objective, 0.125 NA 4/8/2013 Data for 0.4861 to 0.6563 µm. Surface: Image MDE_04_OTS.zmx Configuration 1 of 1 0.4861 0.5876 0.6563 OBJ: 5.0000 mm IMA: 0.000 mm IMA: -0.508 mm 40.00 OBJ: 0.0000 mm Surface: IMA Spot Diagram Stock lens 10x microscope objective, 0.125 NA 4/8/2013 Units are µm. Field : 1 2 RMS radius : 2.132 4.439 GEO radius : 4.090 12.831 Scale bar : 40 Reference : Chief Ray MDE_04_OTS.zmx Configuration 1 of 1 pg. 18 ECEN 4616/5616 1 Spherical 4/8/2013 2 3 Coma 4 5 6 Astigmatism 7 Field Curvature 8 STO Distortion 10 Axial Color 11 SUM Lateral Color Seidel Diagram Stock lens 10x microscope objective, 0.125 NA 4/8/2013 Wavelength: 0.5876 µm. Maximum aberration scale is 0.01000 Millimeters. Grid lines are spaced 0.00100 Millimeters. MDE_04_OTS.zmx Configuration 1 of 1 pg. 19