Table 1.

Imaging the whisker barrel local circuit. Estimates of requirements to acquire and analyze a data volumes of 0.6 × 0.6 × 1.2 um, contain one complete whisker barrel. The 1 × 1 × 5 mm block is certain to contain at least one complete whisker barrel, and parts of several more. Note that array fabrication and staining steps require only a very small fraction of the total process time, with the automated image acquisition process (steps 4 and 6), requiring the bulk of the time. Note also that the image acquisition rate can be scaled up linearly by using multiple automated microscopes in parallel on separate array slides.

Task	Specifications	Time Requirement
1. Specimen Preparation	Fix, Embed Mouse Whisker Barrel	24 Hrs
	Trim, Mount 1 × 1 × 1.5 mm Block	1 Hr
2. Array Preparation	Cut 20,000 Ultrathin (50 nm) Sections	56 Hrs
	Mount Ribbons on 24 50×75mm Slides	24 Hrs
	Map 24 Slides	24 Hrs
3. Array Staining	Stain 24 Slides (for LM and EM cycles)	96 Hrs
4. Fluorescence Acquisition	100 nm pixels (avg. exposure 250 msec/frame) 36 Immunofluorescence channels (9 cycles) 18 fields (2K×2K @ 63x) tile each section	1620 Hrs (= 68 Days) (7 Days on 10 ‘Scopes)
5. Fluorescence Analysis	36,000 cycles/voxel on 1000 1GHz pipelines 36 Channels × 0.86 Teravoxels = 31 Terabytes	8.6 Hrs
6. BsSEM Acquisition	12.5 nm pixels (1 MHz pixel acquisition rate) 288 fields (4K×4K) tile each section	15,360 Hrs (= 640 Days) (64 Days on 10 ‘Scopes)
7. BsSEM Analysis	(10,000 cycles/voxel on 1000 1GHz pipelines) 55 Teravoxels = 55 Terabytes	154 Hrs